Aryl- or heteroaryl-substituted benzene compounds

ABSTRACT

The present invention relates to aryl- or heteroaryl-substituted benzene compounds. The present invention also relates to pharmaceutical compositions containing these compounds and methods of treating cancer by administering these compounds and pharmaceutical compositions to subjects in need thereof. The present invention also relates to the use of such compounds for research or other non-therapeutic purposes.

RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 14/876,658, filed Oct. 6, 2015 (now allowed), which is acontinuation application of U.S. application Ser. No. 14/742,481, filedJun. 17, 2015 (now allowed), which is a continuation application of U.S.application Ser. No. 14/275,667, filed May 12, 2014, now U.S. Pat. No.9,090,562, which is a continuation application of U.S. application Ser.No. 13/722,807, filed Dec. 20, 2012, now U.S. Pat. No. 8,765,732, whichis a continuation application of U.S. application Ser. No. 13/447,007,filed Apr. 13, 2012, now U.S. Pat. No. 8,410,088, which claims priorityto, and the benefit of, U.S. provisional application Nos. 61/474,821,filed Apr. 13, 2011, and 61/499,595 filed Jun. 21, 2011. The entirecontents of each of these applications are incorporated herein byreference in their entireties.

INCORPORATION-BY-REFERENCE OF SEQUENCE LISTING

The contents of the text file named “41478_507001WOST25.txt,” which wascreated on Mar. 28, 2012 and is 2 KB in size, are hereby incorporated byreference in their entireties.

BACKGROUND OF THE INVENTION

In eukaryotic cells DNA is packaged with histones to form chromatin.Changes in the ordered structure of chromatin can lead to alterations intranscription of associated genes. Control of changes in chromatinstructure (and hence of transcription) is mediated by covalentmodifications to histones, most notably of their N-terminal tails. Thesemodifications are often referred to as epigenetic because they can leadto heritable changes in gene expression, but do not affect the sequenceof the DNA itself. Covalent modifications (for example, methylation,acetylation, phosphorylation, and ubiquitination) of the side chains ofamino acids are enzymatically mediated. The selective addition of methylgroups to specific amino acid sites on histones is controlled by theaction of a unique family of enzymes known as histone methyltransferases(HMTs).

The orchestrated collection of biochemical systems behindtranscriptional regulation must be tightly controlled in order for cellgrowth and differentiation to proceed optimally. Disease states resultwhen these controls are disrupted by aberrant expression and/or activityof the enzymes responsible for DNA and histone modification. In humancancers, for example, there is a growing body of evidence to suggestthat dysregulated epigenetic enzyme activity contributes to theuncontrolled cell proliferation associated with cancer as well as othercancer-relevant phenotypes such as enhanced cell migration and invasion.Beyond cancer, there is growing evidence for a role of epigeneticenzymes in a number of other human diseases, including metabolicdiseases (such as diabetes), inflammatory diseases (such as Crohn'sdisease), neurodegenerative diseases (such as Alzheimer's disease) andcardiovascular diseases. Therefore, selectively modulating the aberrantaction of epigenetic enzymes may hold promise for the treatment of arange of diseases.

Polycomb group (PcG) and trithorax group (trxG) proteins are known to bepart of the cellular memory system. See, e.g., Francis et al. (2001) NatRev Mol Cell Biol 2:409-21 and Simon et al. (2002) Curr Opin Genet Dev12:210-8. In general, PcG proteins are transcriptional repressors thatmaintain the “off state,” and trxG proteins are transcriptionalactivators that maintain the “on state.” Because members of PcG and trxGproteins contain intrinsic histone methyltransferase (HMTase) activity,PcG and trxG proteins may participate in cellular memory throughmethylation of core histones. See, e.g., Beisel et al. (2002) Nature419:857-62; Cao et al. (2002) Science 298:1039-43; Czermin et al. (2002)Cell 111:185-96; Kuzmichev et al. (2002) Genes Dev 16:2893-905; Milne etal. (2002) Mol Cell 10:1107-17; Muller et al. (2002) Cell 111:197-208;and Nakamura et al. (2002) Mol Cell 10:1119-28.

Biochemical and genetic studies have provided evidence that DrosophilaPcG proteins function in at least two distinct protein complexes, thePolycomb repressive complex 1 (PRC1) and the ESC-E(Z) complex (alsoknown as Polycomb repressive complex 2 (PRC2)). Otte et al. (2003) CurrOpin Genet Dev 13:448-54. Studies in Drosophila have demonstrated thatthe ESC-E(Z)/EED-EZH2 (i.e., PRC2) complexes have intrinsic histonemethyltransferase activity. Although the compositions of the complexesisolated by different groups are slightly different, they generallycontain EED, EZH2, SUZ12, and RbAp48 or Drosophila homologs thereof.However, a reconstituted complex comprising only EED, EZH2, and SUZ12retains histone methyltransferase activity for lysine 27 of histone H3.U.S. Pat. No. 7,563,589.

Of the various proteins making up PRC2 complexes, EZH2 (Enhancer ofZeste Homolog 2) is the catalytic subunit. The catalytic site of EZH2 inturn is present within a SET domain, a highly conserved sequence motif(named after Su(var)3-9, Enhancer of Zeste, Trithorax) that is found inseveral chromatin-associated proteins, including members of both theTrithorax group and Polycomb group. SET domain is characteristic of allknown histone lysine methyltransferases except the H3-K79methyltransferase DOT 1.

In addition to Hox gene silencing, PRC2-mediated histone H3-K27methylation has been shown to participate in X-inactivation. Plath etal. (2003) Science 300:131-5; Silva et al. (2003) Dev Cell 4:481-95.Recruitment of the PRC2 complex to X₁ and subsequent trimethylation onhistone H3-K27 occurs during the initiation stage of X-inactivation andis dependent on Xist RNA. Furthermore, EZH2 and its associated histoneH3-K27 methyltransferase activity were found to mark differentially thepluripotent epiblast cells and the differentiated trophectoderm, andconsistent with a role of EZH2 in maintaining the epigeneticmodification patterns of pluripotent epiblast cells, Cre-mediateddeletion of EZH2 results in loss of histone H3-K27 methylation in thecells. Erhardt et al. (2003) Development 130:4235-48). Further, studiesin prostate and breast cancer cell lines and tissues have revealed astrong correlation between the levels of EZH2 and SUZ12 and theinvasiveness of these cancers, indicating that dysfunction of the PRC2complex may contribute to cancer. Bracken et al. (2003) EMBOJ22:5323-35; Kirmizis et al. (2003) Mol Cancer Ther 2:113-21; Kleer etal. (2003) Proc Natl Acad Sci USA 100:11606-11; Varambally et al. (2002)Nature 419:624-9.

Recently, somatic mutations of tyrosine 641 (Y641C, Y641F, Y641N, Y641Sand Y641H, sometimes also referred to as Y646C, Y646F, Y646N, Y646S andY646H, respectively) of EZH2 were reported to be associated withfollicular lymphoma (FL) and the germinal center B cell-like (GCB)subtype of diffuse large B-cell lymphoma (DLBCL). Morin et al. (2010)Nat Genet 42:181-5. In all cases, occurrence of the mutant EZH2 gene wasfound to be heterozygous, and expression of both wild-type and mutantalleles was detected in the mutant samples profiled by transcriptomesequencing. It was also demonstrated that all of the mutant forms ofEZH2 could be incorporated into the multi-protein PRC2 complex, but thatthe resulting complexes lacked the ability to catalyze methylation ofthe H3-K27 equivalent residue of a peptidic substrate. Hence, it wasconcluded that the disease-associated changes at Tyr641 of EZH2 resultedin loss of function with respect to EZH2-catalyzed H3-K27 methylation.

SUMMARY OF THE INVENTION

In one aspect, the present invention features an aryl- orheteroaryl-substituted benzene compound of Formula (I) below or apharmaceutically acceptable salt or ester thereof.

In this formula,

-   -   X₁ is N or CR₁₁;    -   X₂ is N or CR₁₃;    -   Z is NR₇R₈, OR₇, S(O)_(n)R₇, or CR₇R₈R₁₄, in which n is 0, 1, or        2;    -   each of R₁, R₅, R₉, and R₁₀, independently, is H or C₁-C₆ alkyl        optionally substituted with one or more substituents selected        from the group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆        alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   each of R₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₁ is H, halo, hydroxyl,        COOH, cyano, or R_(S1), in which R_(S1) is C₁-C₃ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl, C(O)O—C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, 4 to 12-membered heterocycloalkyl, or 5- or        6-membered heteroaryl, and R_(S1) is optionally substituted with        one or more substituents selected from the group consisting of        halo, hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   R₆ is C₆-C₁₀ aryl or 5- or 6-membered heteroaryl, each of which        is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano,        —OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, —C(O)R_(a),        —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —NR_(b)C(O)OR_(a), —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in        which each of R_(a), R_(b), and R_(c), independently is H or        R_(S3), A⁻ is a pharmaceutically acceptable anion, each of        R_(S2) and R_(S3), independently, is C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, or        5- or 6-membered heteroaryl, or R_(a) and R_(b), together with        the N atom to which they are attached, form a 4 to 12-membered        heterocycloalkyl ring having 0 or 1 additional heteroatom, and        each of R_(S2), R_(S3), and the 4 to 12-membered        heterocycloalkyl ring formed by R_(a) and R_(b), is optionally        substituted with one or more one or more -Q₃-T₃, wherein Q₃ is a        bond or C₁-C₃ alkyl linker each optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from        the group consisting of halo, cyano, C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, 5-        or 6-membered heteroaryl, OR_(d), COOR_(d), —S(O)₂R_(d),        —NR_(d)R_(e), and —C(O)NR_(d)R_(e), each of R_(d) and R_(e)        independently being H or C₁-C₆ alkyl, or -Q₃-T₃ is oxo; or any        two neighboring -Q₂-T₂, together with the atoms to which they        are attached form a 5- or 6-membered ring optionally containing        1-4 heteroatoms selected from N, O and S and optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   R₇ is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄ alkyl linker, or        C₂-C₄ alkenyl linker, each linker optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄ is H, halo, cyano,        NR_(f)R_(g), —OR_(f), —C(O)R_(f), —C(O)OR_(f), —C(O)NR_(f)R_(g),        —C(O)NR_(f)OR_(g), —NR_(f)C(O)R_(g), —S(O)₂R_(f), or R_(S4), in        which each of R_(f) and R_(g), independently is H or R_(S5),        each of R_(S4) and R_(S5), independently is C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,        and each of R_(S4) and R_(S5) is optionally substituted with one        or more -Q₅-T₅, wherein Q₅ is a bond, C(O), C(O)NR_(k),        NR_(k)C(O), S(O)₂, or C₁-C₃ alkyl linker, R_(k) being H or C₁-C₆        alkyl, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, 5- or 6-membered heteroaryl, or S(O)_(q)R_(q)        in which q is 0, 1, or 2 and R_(q) is C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,        and T₅ is optionally substituted with one or more substituents        selected from the group consisting of halo, C₁-C₆ alkyl,        hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl        except when T₅ is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo;    -   each of R₈, R₁₁, R₁₂, and R₁₃, independently, is H, halo,        hydroxyl, COOH, cyano, R_(S6), OR_(S6), or COOR_(S6), in which        R_(S6) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈        cycloalkyl, 4 to 12-membered heterocycloalkyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S6) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino; or R₇ and R₈, together with the N atom to which they        are attached, form a 4 to 11-membered heterocycloalkyl ring        having 0 to 2 additional heteroatoms, or R₇ and R₈, together        with the C atom to which they are attached, form C₃-C₈        cycloalkyl or a 4 to 11-membered heterocycloalkyl ring having 1        to 3 heteroatoms, and each of the 4 to 11-membered        heterocycloalkyl rings or C₃-C₈ cycloalkyl formed by R₇ and R₈        is optionally substituted with one or more -Q₆-T₆, wherein Q₆ is        a bond, C(O), C(O)NR_(m), NR_(m)C(O), S(O)₂, or C₁-C₃ alkyl        linker, R_(m) being H or C₁-C₆ alkyl, and T₆ is H, halo, C₁-C₆        alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆        alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl,        4 to 12-membered heterocycloalkyl, 5- or 6-membered heteroaryl,        or S(O)_(p)R_(p) in which p is 0, 1, or 2 and R_(p) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 12-membered heterocycloalkyl, or 5- or 6-membered        heteroaryl, and T₆ is optionally substituted with one or more        substituents selected from the group consisting of halo, C₁-C₆        alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆        alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl,        4 to 12-membered heterocycloalkyl, and 5- or 6-membered        heteroaryl except when T₆ is H, halo, hydroxyl, or cyano; or        -Q₆-T₆ is oxo; and    -   R₁₄ is absent, H, or C₁-C₆ alkyl optionally substituted with one        or more substituents selected from the group consisting of halo,        hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,        mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, and 5- or        6-membered heteroaryl.

One subset of the compounds of Formula (I) includes those of Formula(Ia):

Another subset of the compounds of Formula (I) includes those of Formula(Ib), (Ic), or (Id):

Still another subset of the compounds of formula (I) includes those ofFormula (Ie), (Ig), (II), or (IIa):

The compounds of Formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (Ig), (II)or (IIa) can include one or more of the following features:

X₁ is CR₁₁ and X₂ is CR₁₃.

X₁ is CR₁₁ and X₂ is N.

X₁ is N and X₂ is CR₁₃.

X₁ is N and X₂ is N.

Z is NR₇R₈.

Z is CR₇R₈R₁₄.

Z is OR₇.

Z is S(O)_(n)R₇, in which n is 0, 1, or 2.

R₆ is unsubstituted C₆-C₁₀ aryl or unsubstituted 5- or 6-memberedheteroaryl.

R₆ is C₆-C₁₀ aryl substituted with one or more -Q₂-T₂ or 5- or6-membered heteroaryl substituted with one or more -Q₂-T₂.

R₆ is phenyl substituted with one or more -Q₂-T₂.

R₆ is 5- or 6-membered heteroaryl containing 1-3 additional heteroatomsselected from N, O, and S and optionally substituted with one or more-Q₂-T₂.

R₆ is quinolinyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl,isothiazolyl, furyl, or thienyl, each of which is optionally substitutedwith one or more -Q₂-T₂.

T₂ is C₁-C₆ alkyl, C₆-C₁₀ aryl, halo, —OR_(a), —NR_(a)R_(b),—(NR_(a)R_(b)R_(c))⁺A⁻, —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b),—NR_(b)C(O)R_(a), —NR_(b)C(O)OR_(a), —S(O)₂R_(a), or —S(O)₂NR_(a)R_(b).

T₂ is —NR_(a)R_(b), in which each of R_(a) and R_(b), independently is Hor C₁-C₆ alkyl, or R_(a) and R_(b), together with the N atom to whichthey are attached, form a 4 to 7-membered heterocycloalkyl ring having 0or 1 additional heteroatom, the C₁-C₆ alkyl and the 4 to 12-membered(e.g., 4 to 7-membered) heterocycloalkyl ring being optionallysubstituted with one or more -Q₃-T₃.

Q₂ is C₁-C₃ alkyl linker optionally substituted with halo or hydroxyl.

Q₂ is a bond or methyl or ethyl linker and T₂ is H, halo, —OR_(a),—NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, or —S(O)₂NR_(a)R_(b).

R₇ is not H.

R₇ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl or 4 to 12-membered (e.g., 4 to7-membered) heterocycloalkyl, each optionally substituted with one ormore -Q₅-T₅.

R₇ is 4 to 7-membered heterocycloalkyl optionally substituted with oneor more -Q₅-T₅.

R₇ is piperidinyl, tetrahydropyran, cyclopentyl, or cyclohexyl, eachoptionally substituted with one -Q₅-T₅.

T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈ cycloalkyl, C₆-C₁₀aryl, or 4 to 12-membered (e.g., 4 to 7-membered) heterocycloalkyl.

Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4 to12-membered (e.g., 4 to 7-membered) heterocycloalkyl.

Q₅ is CO, S(O)₂, or NHC(O); and T₅ is C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, or 4 to 12-membered (e.g., 4 to 7-membered)heterocycloalkyl.

Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

Q₅ is C₁-C₃ alkyl linker and T₅ is C₃-C₈ cycloalkyl, 4 to 7-memberedheterocycloalkyl, or S(O)_(q)R_(q).

R₁₁ is H.

R₇ is cyclopentyl or cyclohexyl, each optionally substituted with one-Q₅-T₅.

Q₅ is NHC(O) and T₅ is C₁-C₆ alkyl or C₁-C₆ alkoxy.

R₇ is isopropyl.

Each of R₂ and R₄, independently is H or C₁-C₆ alkyl optionallysubstituted with amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, orC₆-C₁₀ aryl.

Each of R₂ and R₄ is methyl.

R₁ is H.

R₁₂ is H, methyl, ethyl, ethenyl, or halo.

R₁₂ is methyl.

R₁₂ is ethyl.

R₁₂ is ethenyl.

R₈ is H, methyl, or ethyl.

R₈ is methyl.

R₈ is ethyl.

R₈ is 4 to 7-heterocycloalkyl, e.g., tetrahydropyran.

Z is NR₇R₈ or CR₇R₈R₁₄ wherein R₇ and R₈, together with the atom towhich they are attached, form a ring selected from the group consistingof piperidinyl, morpholinyl, piperazinyl, and cyclohexenyl, eachoptionally substituted with one -Q₆-T₆.

R₁₃ is H or methyl.

R₁₃ is H.

R₃ is H.

A⁻ is Br⁻ or Cl⁻.

The present invention also provides pharmaceutical compositionscomprising one or more pharmaceutically acceptable carriers and one ormore compounds selected from those of any Formula disclosed herein.

Another aspect of this invention is a method of treating or preventingcancer. The method includes administering to a subject in need thereof atherapeutically effective amount of one or more compounds selected fromthose of any Formula disclosed herein.

Unless otherwise stated, any description of a method of treatmentincludes uses of the compounds to provide such treatment or prophylaxisas is described in the specification, as well as uses of the compoundsto prepare a medicament to treat or prevent such condition. Thetreatment includes treatment of human or non-human animals includingrodents and other disease models.

For example, the method comprises the step of administering to a subjecthaving a cancer with aberrant H3-K27 methylation an effective amount ofone or more compounds of any of the Formulae disclosed herein, whereinthe compound(s) inhibits histone methyltransferase activity of EZH2,thereby treating the cancer. Examples of aberrant H3-K27 methylation mayinclude a global increase in and/or altered distribution of H3-K27 di ortri-methylation within the cancer cell chromatin.

For example, the cancer is selected from the group consisting of cancersthat overexpress EZH2 or other PRC2 subunits, contain loss-of-functionmutations in H3-K27 demethylases such as UTX, or overexpress accessoryproteins such as PHF19/PCL3 capable of increasing and or mislocalizingEZH2 activity (see references in Sneeringer et al. Proc Natl Acad SciUSA 107(49):20980-5, 2010).

For example, the method comprises the step of administering to a subjecthaving a cancer overexpressing EZH2 a therapeutically effective amountof one or more compounds of any of the Formulae disclosed herein,wherein the compound(s) inhibits histone methyltransferase activity ofEZH2, thereby treating the cancer.

For example, the method comprises the step of administering to a subjecthaving a cancer with a loss-of-function mutation in the H3-K27demethylase UTX a therapeutically effective amount of one or morecompounds of any Formula disclosed herein, wherein the compound(s)inhibits histone methyltransferase activity of EZH2, thereby treatingthe cancer.

For example, the method comprises the step of administering to a subjecthaving a cancer overexpressing an accessory component(s) of the PRC2,such as PHF19/PCL3, a therapeutically effective amount of one or morecompounds of any Formula disclosed herein, wherein the compound(s)inhibits histone methyltransferase activity of EZH2, thereby treatingthe cancer.

In still another aspect, this invention relates to a method ofmodulating the activity of the wild-type EZH2, the catalytic subunit ofthe PRC2 complex which catalyzes the mono-through tri-methylation oflysine 27 on histone H3 (H3-K27). For example, the present inventionrelates to a method of inhibiting the activity of EZH2 in a cell. Thismethod can be conducted either in vitro or in vivo.

In yet another aspect, this invention features to a method of inhibitingin a subject conversion of H3-K27 to trimethylated H3-K27. The methodcomprises administering to a subject a therapeutically effective amountof one or more of the compound of any of the Formulae disclosed hereinto inhibit histone methyltransferase activity of EZH2, therebyinhibiting conversion of H3-K27 to trimethylated H3-K27 in the subject.

For example, the method comprises the step of administering to a subjecthaving a cancer expressing a Y641 mutant of EZH2 a therapeuticallyeffective amount of one or more compounds of any Formula disclosedherein, wherein the compound(s) inhibits histone methyltransferaseactivity of EZH2, thereby treating the cancer.

For example, the cancer is selected from the group consisting offollicular lymphoma and diffuse large B-cell lymphoma (DLBCL) ofgerminal center B cell-like (GCB) subtype. For example, the cancer islymphoma, leukemia or melanoma. Preferably, the lymphoma is non-Hodgkinlymphoma, follicular lymphoma or diffuse large B-cell lymphoma.Alternatively, the leukemia is chronic myelogenous leukemia (CML), acutemyeloid leukemia, acute lymphocytic leukemia or mixed lineage leukemia.

The precancerous condition is myelodysplastic syndromes (MDS, formerlyknown as preleukemia).

For example, the cancer is a hematological cancer.

For example, the method comprises the step of administering to a subjecthaving a cancer expressing a Y641 mutant of EZH2 a therapeuticallyeffective amount of one or more compounds of any Formulae disclosedherein, wherein the compound(s) selectively inhibits histonemethyltransferase activity of the Y641 mutant of EZH2, thereby treatingthe cancer.

For example, the method further comprises the steps of performing anassay to detect a Y641 mutant of EZH2 in a sample comprising cancercells from a subject having a cancer.

In still another aspect, this invention relates to a method ofmodulating the activity of the wild-type and mutant histonemethyltransferase EZH2, the catalytic subunit of the PRC2 complex whichcatalyzes the mono- through tri-methylation of lysine 27 on histone H3(H3-K27). For example, the present invention relates to a method ofinhibiting the activity of certain mutant forms of EZH2 in a cell. Themutant forms of EZH2 include a substitution of another amino acidresidue for tyrosine 641 (Y641, also Tyr641) of wild-type EZH2. Themethod includes contacting the cell with an effective amount of one ormore of the compound of any of Formulae disclosed herein. This methodcan be conducted either in vitro or in vivo.

In yet another aspect, this invention features to a method of inhibitingin a subject conversion of H3-K27 to trimethylated H3-K27. The methodcomprises administering to a subject expressing a Y641 mutant of EZH2 atherapeutically effective amount of one or more of the compound of anyof Formulae disclosed herein to inhibit histone methyltransferaseactivity of EZH2, thereby inhibiting conversion of H3-K27 totrimethylated H3-K27 in the subject. For example, the histonemethyltransferase activity inhibited is that of the Y641 mutant of EZH2.For example, the compound of this invention selectively inhibits histonemethyltransferase activity of the Y641 mutant of EZH2. For example, theY641 mutant of EZH2 is selected from the group consisting of Y641C,Y641F, Y641H, Y641N, and Y641S.

The method of inhibiting in a subject conversion of H3-K27 totrimethylated H3-K27 may also comprise performing an assay to detect aY641 mutant of EZH2 in a sample from a subject before administering tothe subject expressing a Y641 mutant of EZH2 a therapeutically effectiveamount of one or more of the compound of any of Formulae disclosedherein. For example, performing the assay to detect the Y641 mutant ofEZH2 includes whole-genome resequencing or target region resequencingthat detects a nucleic acid encoding the Y641 mutant of EZH2. Forexample, performing the assay to detect the Y641 mutant of EZH2 includescontacting the sample with an antibody that binds specifically to apolypeptide or fragment thereof characteristic of the Y641 mutant ofEZH2. For example, performing the assay to detect the Y641 mutant ofEZH2 includes contacting the sample under highly stringent conditionswith a nucleic acid probe that hybridizes to a nucleic acid encoding apolypeptide or fragment thereof characteristic of the Y641 mutant ofEZH2.

Further, the invention also relates to a method of identifying aninhibitor of a Y641 mutant of EZH2. The method comprises the steps ofcombining an isolated Y641 mutant of EZH2 with a histone substrate, amethyl group donor, and a test compound, wherein the histone substratecomprises a form of H3-K27 selected from the group consisting ofunmethylated H3-K27, monomethylated H3-K27, dimethylated H3-K27, and anycombination thereof; and performing an assay to detect methylation ofH3-K27 (e.g., formation of trimethylated H3-K27) in the histonesubstrate, thereby identifying the test compound as an inhibitor of theY641 mutant of EZH2 when methylation of H3-K27 (e.g., formation oftrimethylated H3-K27) in the presence of the test compound is less thanmethylation of H3-K27 (e.g., formation of trimethylated H3-K27) in theabsence of the test compound.

In one embodiment, performing the assay to detect methylation of H3-K27in the histone substrate comprises measuring incorporation of labeledmethyl groups.

In one embodiment, the labeled methyl groups are isotopically labeledmethyl groups.

In one embodiment, performing the assay to detect methylation of H3-K27in the histone substrate comprises contacting the histone substrate withan antibody that binds specifically to trimethylated H3-K27.

Also within the scope of the invention is a method of identifying aselective inhibitor of a Y641 mutant of EZH2. The method comprises thesteps of combining an isolated Y641 mutant of EZH2 with a histonesubstrate, a methyl group donor, and a test compound, wherein thehistone substrate comprises a form of H3-K27 selected from the groupconsisting of monomethylated H3-K27, dimethylated H3-K27, and acombination of monomethylated H3-K27 and dimethylated H3-K27, therebyforming a test mixture; combining an isolated wild-type EZH2 with ahistone substrate, a methyl group donor, and a test compound, whereinthe histone substrate comprises a form of H3-K27 selected from the groupconsisting of monomethylated H3-K27, dimethylated H3-K27, and acombination of monomethylated H3-K27 and dimethylated H3-K27, therebyforming a control mixture; performing an assay to detect trimethylationof the histone substrate in each of the test mixture and the controlmixture; calculating the ratio of (a) trimethylation with the Y641mutant of EZH2 and the test compound (M+) to (b) trimethylation with theY641 mutant of EZH2 without the test compound (M−); calculating theratio of (c) trimethylation with wild-type EZH2 and the test compound(WT+) to (d) trimethylation with wild-type EZH2 without the testcompound (WT−); comparing the ratio (a)/(b) with the ratio (c)/(d); andidentifying the test compound as a selective inhibitor of the Y641mutant of EZH2 when the ratio (a)/(b) is less than the ratio (c)/(d).

The present invention further provides a method of identifying a subjectas a candidate for treatment with one or more compounds of theinvention. The method comprises the steps of performing an assay todetect a Y641 mutant of EZH2 in a sample from a subject; and identifyinga subject expressing a Y641 mutant of EZH2 as a candidate for treatmentwith one or more compounds of the invention, wherein the compound(s)inhibits histone methyltransferase activity of EZH2.

Still another aspect of the invention is a method of inhibitingconversion of H3-K27 to trimethylated H3-K27. The method comprises thestep of contacting a Y641 mutant of EZH2 with a histone substratecomprising H3-K27 and an effective amount of a compound of the presentinvention, wherein the compound inhibits histone methyltransferaseactivity of EZH2, thereby inhibiting conversion of H3-K27 totrimethylated H3-K27.

Further, the compounds or methods described herein can be used forresearch (e.g., studying epigenetic enzymes) and other non-therapeuticpurposes.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. In the specification, thesingular forms also include the plural unless the context clearlydictates otherwise. Although methods and materials similar or equivalentto those described herein can be used in the practice or testing of thepresent invention, suitable methods and materials are described below.All publications, patent applications, patents and other referencesmentioned herein are incorporated by reference. The references citedherein are not admitted to be prior art to the claimed invention. In thecase of conflict, the present specification, including definitions, willcontrol. In addition, the materials, methods and examples areillustrative only and are not intended to be limiting.

Other features and advantages of the invention will be apparent from thefollowing detailed description and claims.

BRIEF DESCRIPTIONS OF FIGURES

FIG. 1 (A) is an idealized plot of cell count (i.e., cell number) as afunction of time showing exponential proliferation during log-phase cellgrowth.

FIG. 1(B) is an idealized plot of ln(cell count) as a function of timefor the data from panel (A).

FIG. 2 is a graph showing biphasic cell growth curves in the presence ofan antiproliferative compound for which there is a delay before theimpact of the compound on cell growth is realized. The compound beginsto affect cell growth at the time point labeled “start of impact.” Thesolid circles represent idealized data for the vehicle (or solvent)control sample that is not treated with compound. The other symbolsrepresent biphasic growth curves for cells treated with differentconcentrations of compound (i.e., drug).

FIG. 3 is a replot of k_(p) as a function of compound concentration for(A) a cytostatic and (B) a cytotoxic compound, illustrating the graphicdetermination of the LCC for a cytotoxic agent. Note that for acytostatic compound (panel A), the value of k_(p) can never drop belowzero.

FIG. 4 is a diagram showing global H3K27me3 methylation in WSU-DLCL2tumors from mice treated with Compound 87 for 7 days.

FIG. 5 is a diagram showing global H3K27me3 methylation in WSU-DLCL2tumors from mice treated with Compound 141 for 7 days.

FIG. 6 is a diagram showing tumor growth of WSU-DLCL2 xenograft bearingmice over the treatment course of 28 days treated with vehicle orCompound 141.

FIG. 7 is a diagram showing tumor growth of WSU-DLCL2 xenograft bearingmice treated with Compound 44.

FIG. 8 is a diagram showing global H3K27me3 methylation in WSU-DLCL2tumors from mice treated with Compound 44 for 28 and 7 days.

FIG. 9 is a diagram showing tumor growth of WSU-DLCL2 xenograft bearingmice with Compound 44 treatment at different dosing schedules.

FIG. 10 is a diagram showing global H3K27me3 methylation in WSU-DLCL2tumors from mice treated with Compound 44 at different dosing schedulesfor 28 days.

FIG. 11 is a diagram showing effect of Compound 44 on mouse body weight.Data represent the mean+SD (n=9). Dosages which resulted in mortalitiesare not plotted.

FIG. 12 is a diagram showing antitumor effects of orally administeredCompound 44 against a diffuse large B cell lymphoma KARPAS-422 xenograftin mice. Data represent the mean+SD (n=9). * P<0.05 versus vehiclecontrol on day 29 (repeated measures ANOVA followed by Dunnett-typemultiple comparison test). Dosages which resulted in mortalities are notplotted.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel aryl- or heteroaryl-substitutedbenzene compounds, synthetic methods for making the compounds,pharmaceutical compositions containing them and various uses of thecompounds.

1. ARYL- OR HETEROARYL-SUBSTITUTED BENZENE COMPOUNDS

The present invention provides the compounds of Formula (I):

or a pharmaceutically acceptable salt or ester thereof. In this formula:

-   -   X₁ is N or CR₁₁;    -   X₂ is N or CR₁₃;    -   Z is NR₇R₈, OR₇, S(O)_(n)R₇, or CR₇R₈R₁₄, in which n is 0, 1, or        2;    -   each of R₁, R₅, R₉, and R₁₀, independently, is H or C₁-C₆ alkyl        optionally substituted with one or more substituents selected        from the group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆        alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   each of R₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₁ is H, halo, hydroxyl,        COOH, cyano, or R_(S1), in which R_(S1) is C₁-C₃ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl, C(O)O—C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, 4 to 12-membered heterocycloalkyl, or 5- or        6-membered heteroaryl, and R_(S1) is optionally substituted with        one or more substituents selected from the group consisting of        halo, hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   R₆ is C₆-C₁₀ aryl or 5- or 6-membered heteroaryl, each of which        is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano,        —OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, —C(O)R_(a),        —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —NR_(b)C(O)OR_(a), —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in        which each of R_(a), R_(b), and R_(c), independently is H or        R_(S3), A⁻ is a pharmaceutically acceptable anion, each of        R_(S2) and R_(S3), independently, is C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, or        5- or 6-membered heteroaryl, or R_(a) and R_(b), together with        the N atom to which they are attached, form a 4 to 12-membered        heterocycloalkyl ring having 0 or 1 additional heteroatom, and        each of R_(S2), R_(S3), and the 4 to 12-membered        heterocycloalkyl ring formed by R_(a) and R_(b), is optionally        substituted with one or more one or more -Q₃-T₃, wherein Q₃ is a        bond or C₁-C₃ alkyl linker each optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from        the group consisting of halo, cyano, C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, 5-        or 6-membered heteroaryl, OR_(d), COOR_(d), —S(O)₂R_(d),        —NR_(d)R_(e), and —C(O)NR_(d)R_(e), each of R_(d) and R_(e)        independently being H or C₁-C₆ alkyl, or -Q₃-T₃ is oxo; or any        two neighboring -Q₂-T₂, together with the atoms to which they        are attached form a 5- or 6-membered ring optionally containing        1-4 heteroatoms selected from N, O and S and optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   R₇ is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄ alkyl linker, or        C₂-C₄ alkenyl linker, each linker optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄ is H, halo, cyano,        NR_(f)R_(g), —OR_(f), —C(O)R_(f), —C(O)OR_(f), —C(O)NR_(f)R_(g),        —C(O)NR_(f)OR_(g), —NR_(f)C(O)R_(g), —S(O)₂R_(f), or R_(S4), in        which each of R_(f) and R_(g), independently is H or R_(S5),        each of R_(S4) and R_(S5), independently is C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,        and each of R_(S4) and R_(S5) is optionally substituted with one        or more -Q₅-T₅, wherein Q₅ is a bond, C(O), C(O)NR_(k),        NR_(k)C(O), S(O)₂, or C₁-C₃ alkyl linker, R_(k) being H or C₁-C₆        alkyl, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, 5- or 6-membered heteroaryl, or S(O)_(q)R_(q)        in which q is 0, 1, or 2 and R_(q) is C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,        and T₅ is optionally substituted with one or more substituents        selected from the group consisting of halo, C₁-C₆ alkyl,        hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl        except when T₅ is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo;    -   each of R₈, R₁₁, R₁₂, and R₁₃, independently, is H, halo,        hydroxyl, COOH, cyano, R_(S6), OR_(S6), or COOR_(S6), in which        R_(S6) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈        cycloalkyl, 4 to 12-membered heterocycloalkyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S6) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino; or R₇ and R₈, together with the N atom to which they        are attached, form a 4 to 11-membered heterocycloalkyl ring        having 0 to 2 additional heteroatoms, or R₇ and R₈, together        with the C atom to which they are attached, form C₃-C₈        cycloalkyl or a 4 to 11-membered heterocycloalkyl ring having 1        to 3 heteroatoms, and each of the 4 to 11-membered        heterocycloalkyl rings or C₃-C₈ cycloalkyl formed by R₇ and R₈        is optionally substituted with one or more -Q₆-T₆, wherein Q₆ is        a bond, C(O), C(O)NR_(m), NR_(m)C(O), S(O)₂, or C₁-C₃ alkyl        linker, R_(m) being H or C₁-C₆ alkyl, and T₆ is H, halo, C₁-C₆        alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆        alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl,        4 to 12-membered heterocycloalkyl, 5- or 6-membered heteroaryl,        or S(O)_(p)R_(p) in which p is 0, 1, or 2 and R_(p) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 12-membered heterocycloalkyl, or 5- or 6-membered        heteroaryl, and T₆ is optionally substituted with one or more        substituents selected from the group consisting of halo, C₁-C₆        alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆        alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl,        4 to 12-membered heterocycloalkyl, and 5- or 6-membered        heteroaryl except when T₆ is H, halo, hydroxyl, or cyano; or        -Q₆-T₆ is oxo; and    -   R₁₄ is absent, H, or C₁-C₆ alkyl optionally substituted with one        or more substituents selected from the group consisting of halo,        hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,        mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, and 5- or        6-membered heteroaryl.

For example, X₁ is CR₁₁ and X₂ is CR₁₃.

For example, X₁ is CR₁₁ and X₂ is N.

For example, X₁ is N and X₂ is CR₁₃.

For example, X₁ is N and X₂ is N.

For example, Z is NR₇R₈.

For example, Z is CR₇R₈R₁₄.

For example, Z is OR₇.

For example, Z is S(O)_(n)R₇, in which n is 0, 1, or 2.

For example, Z is SR₇.

For example, R₆ is unsubstituted C₆-C₁₀ aryl or unsubstituted 5- or6-membered heteroaryl.

For example, R₆ is C₆-C₁₀ aryl substituted with one or more -Q₂-T₂ or 5-or 6-membered heteroaryl substituted with one or more -Q₂-T₂.

For example, R₆ is unsubstituted phenyl.

For example, R₆ is phenyl substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing 1-3 additionalheteroatoms selected from N, O, and S and optionally substituted withone or more -Q₂-T₂.

For example, R₆ is pyridinyl, pyrazolyl, pyrimidinyl, quinolinyl,tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, furyl, orthienyl, each of which is optionally substituted with one or more-Q₂-T₂.

For example, Q₂ is a bond.

For example, Q₂ is an unsubstituted C₁-C₃ alkyl linker.

For example, T₂ is C₁-C₆ alkyl or C₆-C₁₀ aryl, each optionallysubstituted with one or more -Q₃-T₃.

For example, T₂ is an unsubstituted substituted straight chain C₁-C₆ orbranched C₃-C₆ alkyl, including but not limited to, methyl, ethyl,n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl andn-hexyl.

For example, T₂ is phenyl.

For example, T₂ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, T₂ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and thelike) optionally substituted with one or more -Q₃-T₃.

For example, T₂ is —OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻,—C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),—NR_(b)C(O)OR_(a), —S(O)₂R_(a), or —S(O)₂NR_(a)R_(b).

For example, T₂ is —NR_(a)R_(b) or —C(O)NR_(a)R_(b), in which each ofR_(a) and R_(b), independently is H or C₁-C₆ alkyl, or R_(a) and R_(b),together with the N atom to which they are attached, form a 4 to7-membered heterocycloalkyl ring having 0 or 1 additional heteroatom,the C₁-C₆ alkyl and the 4 to 7-membered heterocycloalkyl ring beingoptionally substituted with one or more -Q₃-T₃.

For example, Q₂ is C₁-C₃ alkyl linker optionally substituted with haloor hydroxyl.

For example, Q₂ is a bond or methyl linker and T₂ is H, halo, —OR_(a),—NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, or —S(O)₂NR_(a)R_(b).

For example, each of R_(a), R_(b), and R_(e), independently is H orC₁-C₆ alkyl optionally substituted with one or more -Q₃-T₃.

For example, one of R_(a), R_(b), and R_(c) is H.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and thelike) and the ring is optionally substituted with one or more -Q₃-T₃.

For example, -Q₃-T₃ is oxo.

For example, T₂ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₃-T₃ are oxo.

For example, Q₃ is a bond or unsubstituted or substituted C₁-C₃ alkyllinker.

For example, T₃ is H, halo, 4 to 7-membered heterocycloalkyl, C₁-C₃alkyl, OR_(d), COOR_(d), —S(O)₂R_(d), or —NR_(d)R_(e).

For example, one of R_(d) and R_(e) is H.

For example, R₇ is not H.

For example, R₇ is —C(O)R_(f).

For example, R₇ is —C(O)R_(f), in which R_(f) is C₃-C₈ cycloalkyl.

For example, R₇ is C₆-C₁₀ aryl substituted with one or more -Q₅-T₅.

For example, R₇ is phenyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₁-C₆ alkyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₃-C₈ cycloalkyl optionally substituted with one ormore -Q₅-T₅.

For example, R₇ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,and morpholinyl, and the like) optionally substituted with one or more-Q₅-T₅.

For example, R₇ is 5 to 6-membered heterocycloalkyl optionallysubstituted with one or more -Q₅-T₅.

For example, R₇ is isopropyl.

For example, R₇ is pyrrolidinyl, piperidinyl, tetrahydropyran,cyclopentyl, cyclohexyl, or cycloheptyl, each optionally substitutedwith one -Q₅-T₅.

For example, R₇ is cyclopentyl or cyclohexyl, each optionallysubstituted with one -Q₅-T₅.

For example, R₇ is pyrrolidinyl, piperidinyl, tetrahydropyran,tetrahydro-2H-thiopyranyl, cyclopentyl, cyclohexyl, or cycloheptyl, eachoptionally substituted with one or more -Q₅-T₅.

For example, R₇ is cyclopentyl, cyclohexyl or tetrahydro-2H-thiopyranyl,each optionally substituted with one or more -Q₅-T₅.

For example, one or more -Q₅-T₅ are oxo.

For example, R₇ is 1-oxide-tetrahydro-2H-thiopyranyl or1,1-dioxide-tetrahydro-2H-thiopyranyl.

For example, Q₅ is a bond and T₅ is amino, mono-C₁-C₆ alkylamino, ordi-C₁-C₆ alkylamino.

For example, Q₅ is NHC(O) and T₅ is C₁-C₆ alkyl or C₁-C₆ alkoxy.

For example, -Q₅-T₅ is oxo.

For example, T₄ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₅-T₅ are oxo.

For example, T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₅ is CO, S(O)₂, or NHC(O); and T₅ is C₁-C₆ alkyl, C₁-C₆alkoxyl, C₃-C₈ cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₅ is C₁-C₆ alkyl or C₁-C₆ alkoxyl, each optionallysubstituted with halo, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is C₃-C₈ cycloalkyl, 4 to7-membered heterocycloalkyl, or S(O)_(q)R_(q).

For example, R₁₁ is H.

For example, each of R₂ and R₄, independently, is H or C₁-C₆ alkyloptionally substituted with amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₆-C₁₀ aryl.

For example, each of R₂ and R₄, independently is C₁-C₃ alkyl optionallysubstituted with C₁-C₆ alkoxyl.

For example, each of R₂ and R₄ is methyl.

For example, R₁ is H.

For example, R₁₂ is H, methyl, ethyl, ethenyl, or halo.

For example, R₁₂ is methyl.

For example, R₁₂ is ethyl.

For example, R₁₂ is ethenyl.

For example, R₈ is H, methyl, ethyl, or ethenyl.

For example, R₈ is methyl.

For example, R₈ is ethyl.

For example, R₈ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and thelike).

For example, R₈ is tetrahydropyran.

For example, R₈ is tetrahydropyran and R₇ is -Q₄-T₄, in which Q₄ is abond or C₁-C₄ alkyl linker and T₄ is H, C₁-C₆ alkyl, C₃-C₈ cycloalkyl or4 to 7-membered heterocycloalkyl.

For example, Z is NR₇R or CR₇R₈R₁₄ wherein R₇ and R₈, together with theatom to which they are attached, form a 4 to 11-memberedheterocycloalkyl ring having 1 to 3 heteroatoms (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,and morpholinyl, 1,4-dioxa-8-azaspiro[4.5]decanyl, and the like) orC₃-C₈ cycloalkyl, each optionally substituted with one or more -Q₆-T₆.

For example, the ring formed by R₇ and R₈ is selected from the groupconsisting of azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl,piperazinyl, 1,4-dioxa-8-azaspiro[4.5]decanyl, and cyclohexenyl, eachoptionally substituted with one -Q₆-T₆.

For example, -Q₆-T₆ is oxo.

For example, T₆ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₆ is a bond and T₆ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₆ is CO, S(O)₂, or NHC(O); and T₆ is C₁-C₆ alkyl, C₁-C₆alkoxyl, C₃-C₈ cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₆ is C₁-C₆ alkyl or C₁-C₆ alkoxyl, each optionallysubstituted with halo, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₆ is C₁-C₃ alkyl linker and T₆ is H or C₆-C₁₀ aryl.

For example, Q₆ is C₁-C₃ alkyl linker and T₆ is C₃-C₈ cycloalkyl, 4 to7-membered heterocycloalkyl, or S(O)_(p)R_(p).

For example, each of R_(p) and R_(q), independently, is C₁-C₆ alkyl.

For example, R₁₃ is H or methyl.

For example, R₁₃ is H.

For example, R₃ is H.

For example, A⁻ is Br or Cl⁻.

For example, each of R₅, R₉, and R₁₀ is H.

The present invention provides the compounds of Formula (Ia)

or a pharmaceutically acceptable salt or ester thereof, wherein:

-   -   X₁ is N or CR₁₁;    -   X₂ is N or CR₁₃;    -   Z is NR₇R₈, OR₇, S(O)_(n)R₇, or CR₇R₈R₁₄, in which n is 0, 1, or        2;    -   each of R₁ and R₅, independently, is H or C₁-C₆ alkyl optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   each of R₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₁ is H, halo, hydroxyl,        COOH, cyano, or R_(S1), in which R_(S1) is C₁-C₃ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl, C(O)O—C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, 4 to 12-membered heterocycloalkyl, or 5- or        6-membered heteroaryl, and R_(S1) is optionally substituted with        one or more substituents selected from the group consisting of        halo, hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   R₆ is C₆-C₁₀ aryl or 5- or 6-membered heteroaryl, each of which        is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano,        —OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, —C(O)R_(a),        —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —NR_(b)C(O)OR_(a), —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in        which each of R_(a), R_(b), and R_(c), independently is H or        R_(S3), A⁻ is a pharmaceutically acceptable anion, each of        R_(S2) and R_(S3), independently, is C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, or        5- or 6-membered heteroaryl, or R_(a) and R_(b), together with        the N atom to which they are attached, form a 4 to 12-membered        heterocycloalkyl ring having 0 or 1 additional heteroatom, and        each of R_(S2), R_(S3), and the 4 to 12-membered        heterocycloalkyl ring formed by R_(a) and R_(b), is optionally        substituted with one or more one or more -Q₃-T₃, wherein Q₃ is a        bond or C₁-C₃ alkyl linker each optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from        the group consisting of halo, cyano, C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, 5-        or 6-membered heteroaryl, OR_(d), COOR_(d), —S(O)₂R_(d),        —NR_(d)R_(e), and —C(O)NR_(d)R_(e), each of R_(d) and R_(e)        independently being H or C₁-C₆ alkyl, or -Q₃-T₃ is oxo; or any        two neighboring -Q₂-T₂, together with the atoms to which they        are attached form a 5- or 6-membered ring optionally containing        1-4 heteroatoms selected from N, O and S and optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, and 5- or 6-membered heteroaryl;    -   R₇ is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄ alkyl linker, or        C₂-C₄ alkenyl linker, each linker optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄ is H, halo, cyano,        NR_(f)R_(g), —OR_(f), —C(O)R_(f), —C(O)OR_(f), —C(O)NR_(f)R_(g),        —C(O)NR_(f)OR_(g), —NR_(f)C(O)R_(g), —S(O)₂R_(f), or R_(S4), in        which each of R_(f) and R_(g), independently is H or R_(S5),        each of R_(S4) and R_(S5), independently is C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,        and each of R_(S4) and R_(S5) is optionally substituted with one        or more -Q₅-T₅, wherein Q₅ is a bond, C(O), C(O)NR_(k),        NR_(k)C(O), S(O)₂, or C₁-C₃ alkyl linker, R_(k) being H or C₁-C₆        alkyl, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered        heterocycloalkyl, 5- or 6-membered heteroaryl, or S(O)_(q)R_(q)        in which q is 0, 1, or 2 and R_(q) is C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,        and T₅ is optionally substituted with one or more substituents        selected from the group consisting of halo, C₁-C₆ alkyl,        hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl        except when T₅ is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo;    -   each of R₈, R₁₁, R₁₂, and R₁₃, independently, is H, halo,        hydroxyl, COOH, cyano, R_(S6), OR_(S6), or COOR_(S6), in which        R_(S6) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈        cycloalkyl, 4 to 12-membered heterocycloalkyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S6) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino; or R₇ and R₈, together with the N atom to which they        are attached, form a 4 to 11-membered heterocycloalkyl ring        having 0 to 2 additional heteroatoms, or R₇ and R₈, together        with the C atom to which they are attached, form C3-C8        cycloalkyl or a 4 to 11-membered heterocycloalkyl ring having 1        to 3 heteroatoms, and each of the 4 to 11-membered        heterocycloalkyl rings or C₃-C₈ cycloalkyl formed by R₇ and R₈        is optionally substituted with one or more -Q₆-T₆, wherein Q₆ is        a bond, C(O), C(O)NR_(m), NR_(m)C(O), S(O)₂, or C₁-C₃ alkyl        linker, R_(m) being H or C₁-C₆ alkyl, and T₆ is H, halo, C₁-C₆        alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆        alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl,        4 to 12-membered heterocycloalkyl, 5- or 6-membered heteroaryl,        or S(O)_(p)R_(p) in which p is 0, 1, or 2 and R_(p) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5- or 6-membered        heteroaryl, and T₆ is optionally substituted with one or more        substituents selected from the group consisting of halo, C₁-C₆        alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆        alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl,        4 to 12-membered heterocycloalkyl, and 5- or 6-membered        heteroaryl except when T₆ is H, halo, hydroxyl, or cyano; or        -Q₆-T₆ is oxo; and    -   R₁₄ is absent, H, or C₁-C₆ alkyl optionally substituted with one        or more substituents selected from the group consisting of halo,        hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,        mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, and 5- or        6-membered heteroaryl.

For example, X₂ is CR₁₃.

For example, X₂ is N.

For example, Z is NR₇R₈.

For example, Z is CR₇R₈R₁₄.

For example, Z is OR₇.

For example, Z is S(O)_(n)R₇, in which n is 0, 1, or 2.

For example, Z is SR₇.

For example, R₆ is unsubstituted C₆-C₁₀ aryl or unsubstituted 5- or6-membered heteroaryl.

For example, R₆ is C₆-C₁₀ aryl substituted with one or more -Q₂-T₂ or 5-or 6-membered heteroaryl substituted with one or more -Q₂-T₂.

For example, R₆ is unsubstituted phenyl.

For example, R₆ is phenyl substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing 1-3 additionalheteroatoms selected from N, O, and S and optionally substituted withone or more -Q₂-T₂.

For example, R₆ is pyridinyl, pyrazolyl, pyrimidinyl, quinolinyl,tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, furyl, orthienyl, each of which is optionally substituted with one or more-Q₂-T₂.

For example, Q₂ is a bond.

For example, Q₂ is an unsubstituted C₁-C₃ alkyl linker.

For example, T₂ is C₁-C₆ alkyl or C₆-C₁₀ aryl, each optionallysubstituted with one or more -Q₃-T₃.

For example, T₂ is an unsubstituted substituted straight chain C₁-C₆ orbranched C₃-C₆ alkyl, including but not limited to, methyl, ethyl,n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl andn-hexyl.

For example, T₂ is phenyl.

For example, T₂ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, T₂ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and thelike) optionally substituted with one or more -Q₃-T₃.

For example, T₂ is —OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻,—C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),—NR_(b)C(O)OR_(a), —S(O)₂R_(a), or —S(O)₂NR_(a)R_(b).

For example, T₂ is —NR_(a)R_(b) or —C(O)NR_(a)R_(b), in which each ofR_(a) and R_(b), independently is H or C₁-C₆ alkyl, or R_(a) and R_(b),together with the N atom to which they are attached, form a 4 to7-membered heterocycloalkyl ring having 0 or 1 additional heteroatom,the C₁-C₆ alkyl and the 4 to 7-membered heterocycloalkyl ring beingoptionally substituted with one or more -Q₃-T₃.

For example, Q₂ is C₁-C₃ alkyl linker optionally substituted with haloor hydroxyl.

For example, Q₂ is a bond or methyl linker and T₂ is H, halo, —OR_(a),—NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, or —S(O)₂NR_(a)R_(b).

For example, each of R_(a), R_(b), and R_(e), independently is H orC₁-C₆ alkyl optionally substituted with one or more -Q₃-T₃.

For example, one of R_(a), R_(b), and R_(c) is H.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and thelike) and the ring is optionally substituted with one or more -Q₃-T₃.

For example, -Q₃-T₃ is oxo.

For example, T₂ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₃-T₃ are oxo.

For example, Q₃ is a bond or unsubstituted or substituted C₁-C₃ alkyllinker.

For example, T₃ is H, halo, 4 to 7-membered heterocycloalkyl, C₁-C₃alkyl, OR_(d), COOR_(d), —S(O)₂R_(d), or —NR_(d)R_(e).

For example, one of R_(d) and R_(e) is H.

For example, R₇ is not H.

For example, R₇ is —C(O)R_(f).

For example, R₇ is —C(O)R_(f), in which R_(f) is C₃-C₈ cycloalkyl.

For example, R₇ is C₆-C₁₀ aryl substituted with one or more -Q₅-T₅.

For example, R₇ is phenyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₁-C₆ alkyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₃-C₈ cycloalkyl optionally substituted with one ormore -Q₅-T₅.

For example, R₇ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,and morpholinyl, and the like) optionally substituted with one or more-Q₅-T₅.

For example, R₇ is 5 to 6-membered heterocycloalkyl optionallysubstituted with one or more -Q₅-T₅.

For example, R₇ is isopropyl.

For example, R₇ is pyrrolidinyl, piperidinyl, tetrahydropyran,cyclopentyl, cyclohexyl, or cycloheptyl, each optionally substitutedwith one -Q₅-T₅.

For example, R₇ is cyclopentyl or cyclohexyl, each optionallysubstituted with one -Q₅-T₅.

For example, R₇ is pyrrolidinyl, piperidinyl, tetrahydropyran,tetrahydro-2H-thiopyranyl, cyclopentyl, cyclohexyl, or cycloheptyl, eachoptionally substituted with one or more -Q₅-T₅.

For example, R₇ is cyclopentyl, cyclohexyl or tetrahydro-2H-thiopyranyl,each optionally substituted with one or more -Q₅-T₅.

For example, one or more -Q₅-T₅ are oxo.

For example, R₇ is 1-oxide-tetrahydro-2H-thiopyranyl or1,1-dioxide-tetrahydro-2H-thiopyranyl.

For example, Q₅ is a bond and T₅ is amino, mono-C₁-C₆ alkylamino, ordi-C₁-C₆ alkylamino.

For example, Q₅ is NHC(O) and T₅ is C₁-C₆ alkyl or C₁-C₆ alkoxy.

For example, -Q₅-T₅ is oxo.

For example, T₄ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₅-T₅ are oxo.

For example, T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₅ is CO, S(O)₂, or NHC(O); and T₅ is C₁-C₆ alkyl, C₁-C₆alkoxyl, C₃-C₈ cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₅ is C₁-C₆ alkyl or C₁-C₆ alkoxyl, each optionallysubstituted with halo, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is C₃-C₈ cycloalkyl, 4 to7-membered heterocycloalkyl, or S(O)_(q)R_(q).

For example, R₁₁ is H.

For example, each of R₂ and R₄, independently, is H or C₁-C₆ alkyloptionally substituted with amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₆-C₁₀ aryl.

For example, each of R₂ and R₄, independently is C₁-C₃ alkyl optionallysubstituted with C₁-C₆ alkoxyl.

For example, each of R₂ and R₄ is methyl.

For example, R₁ is H.

For example, R₅ is H.

For example, R₁₂ is H, methyl, ethyl, ethenyl, or halo.

For example, R₁₂ is methyl.

For example, R₁₂ is ethyl.

For example, R₁₂ is ethenyl.

For example, R₈ is H, methyl, ethyl, or ethenyl.

For example, R₈ is methyl.

For example, R₈ is ethyl.

For example, R₈ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and thelike).

For example, R₈ is tetrahydropyran.

For example, R₈ is tetrahydropyran and R₇ is -Q₄-T₄, in which Q₄ is abond or C₁-C₄ alkyl linker and T₄ is H, C₁-C₆ alkyl, C₃-C₈ cycloalkyl or4 to 7-membered heterocycloalkyl.

For example, Z is NR₇R or CR₇R₈R₁₄ wherein R₇ and R₈, together with theatom to which they are attached, form a 4 to 11-memberedheterocycloalkyl ring having 1 to 3 heteroatoms (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,and morpholinyl, 1,4-dioxa-8-azaspiro[4.5]decanyl, and the like) orC₃-C₈ cycloalkyl, each optionally substituted with one or more -Q₆-T₆.

For example, the ring formed by R₇ and R₈ is selected from the groupconsisting of azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl,piperazinyl, 1,4-dioxa-8-azaspiro[4.5]decanyl, and cyclohexenyl, eachoptionally substituted with one -Q₆-T₆.

For example, -Q₆-T₆ is oxo.

For example, T₆ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₆ is a bond and T₆ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₆ is CO, S(O)₂, or NHC(O); and T₆ is C₁-C₆ alkyl, C₁-C₆alkoxyl, C₃-C₈ cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₆ is C₁-C₆ alkyl or C₁-C₆ alkoxyl, each optionallysubstituted with halo, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₆ is C₁-C₃ alkyl linker and T₆ is H or C₆-C₁₀ aryl.

For example, Q₆ is C₁-C₃ alkyl linker and T₆ is C₃-C₈ cycloalkyl, 4 to7-membered heterocycloalkyl, or S(O)_(p)R_(p).

For example, each of R_(p) and R_(q), independently, is C₁-C₆ alkyl.

For example, R₁₃ is H or methyl.

For example, R₁₃ is H.

For example, R₃ is H.

For example, A⁻ is Br or Cl⁻.

The present invention provides the compounds of Formula (Ib), (Ic), or(Id):

or pharmaceutically acceptable salts or esters thereof, wherein Z, X₂,R₁, R₂, R₃, R₄, R₅, R₆, R₁₁, and R₁₂ are defined herein.

Still another subset of the compounds of formula (I) includes those ofFormula (Ie), or (Ig):

or a pharmaceutically acceptable salts or esters thereof, wherein Z, X₂,R₂, R₃, R₄, R₆, and R₁₂ are defined herein.

For example, R₂, R₄ and R₁₂ are each, independently C₁₋₆ alkyl.

For example, R₆ is C₆-C₁₀ aryl or 5- or 6-membered heteroaryl, each ofwhich is optionally, independently substituted with one or more -Q₂-T₂,wherein Q₂ is a bond or C₁-C₃ alkyl linker, and T₂ is H, halo, cyano,—OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, —C(O)NR_(a)R_(b),—NR_(b)C(O)R_(a), —S(O)₂R_(a), or R_(S2), in which each of R_(a) andR_(b), independently is H or R_(S3), each of R_(S2) and R_(S3),independently, is C₁-C₆ alkyl, or R_(a) and R_(b), together with the Natom to which they are attached, form a 4 to 7-membered heterocycloalkylring having 0 or 1 additional heteroatom, and each of R_(S2), R_(S3),and the 4 to 7-membered heterocycloalkyl ring formed by R_(a) and R_(b),is optionally, independently substituted with one or more -Q₃-T₃,wherein Q₃ is a bond or C₁-C₃ alkyl linker and T₃ is selected from thegroup consisting of halo, C₁-C₆ alkyl, 4 to 7-membered heterocycloalkyl,OR_(d), —S(O)₂R_(d), and —NR_(d)R_(e), each of R_(d) and R_(e)independently being H or C₁-C₆ alkyl, or -Q₃-T₃ is oxo; or any twoneighboring -Q₂-T₂, together with the atoms to which they are attachedform a 5- or 6-membered ring optionally containing 1-4 heteroatomsselected from N, O and S.

Another subset of the compounds of Formula (I) includes those of Formula(II):

or a pharmaceutically acceptable salts or esters thereof, wherein

-   -   Q₂ is a bond or methyl linker;    -   T₂ is H, halo, —OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, or        —S(O)₂NR_(a)R_(b);    -   R₇ is piperidinyl, tetrahydropyran, cyclopentyl, or cyclohexyl,        each optionally substituted with one -Q₅-T₅;    -   R₈ is ethyl and    -   R_(a), R_(b), and R_(c) are defined herein.

For example, Q₂ is a bond

For example, Q₂ is a methyl linker

For example, T₂ is —NR_(a)R_(b) or —(NR_(a)R_(b)R_(c))⁺A⁻.

Yet another subset of the compounds of Formula (I) includes those ofFormula (IIa):

or a pharmaceutically acceptable salts or esters thereof, wherein R₇,R₈, R_(a), R_(b), and R_(c) are defined herein.

The compounds of Formula (II) or (IIa) can include one or more of thefollowing features:

For example, each of R_(a) and R_(b), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₃-T₃.

For example, one of R_(a) and R_(b) is H.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl, andthe like) and the ring is optionally substituted with one or more-Q₃-T₃.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl, or morpholinyl,and the ring is optionally substituted with one or more -Q₃-T₃.

For example, one or more -Q₃-T₃ are oxo.

For example, Q₃ is a bond or unsubstituted or substituted C₁-C₃ alkyllinker.

For example, T₃ is H, halo, 4 to 7-membered heterocycloalkyl, C₁-C₃alkyl, OR_(d), COOR_(d), —S(O)₂R_(d), or —NR_(d)R_(e).

For example, one of R_(d) and R_(e) is H.

For example, R₇ is C₃-C₈ cycloalkyl or 4 to 7-membered heterocycloalkyl,each optionally substituted with one or more -Q₅-T₅.

For example, R₇ is piperidinyl, tetrahydropyran,tetrahydro-2H-thiopyranyl, cyclopentyl, cyclohexyl, pyrrolidinyl, orcycloheptyl, each optionally substituted with one or more -Q₅-T₅.

For example, R₇ is cyclopentyl cyclohexyl or tetrahydro-2H-thiopyranyl,each optionally substituted with one or more -Q₅-T₅.

For example, Q₅ is NHC(O) and T₅ is C₁-C₆ alkyl or C₁-C₆ alkoxy.

For example, one or more -Q₅-T₅ are oxo.

For example, R₇ is 1-oxide-tetrahydro-2H-thiopyranyl or1,1-dioxide-tetrahydro-2H-thiopyranyl.

For example, Q₅ is a bond and T₅ is amino, mono-C₁-C₆ alkylamino,di-C₁-C₆ alkylamino.

For example, Q₅ is CO, S(O)₂, or NHC(O); and T₅ is C₁-C₆ alkyl, C₁-C₆alkoxyl, C₃-C₈ cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, R₈ is H or C₁-C₆ alkyl which is optionally substituted withone or more substituents selected from the group consisting of halo,hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,mono-C₁-C₆ alkylamino, and di-C₁-C₆ alkylamino.

For example, R₈ is H, methyl, or ethyl.

Still another subset of compounds of Formula (I) includes those ofFormula (III):

or a pharmaceutically acceptable salts or esters thereof,wherein

R₃ is hydrogen, C₁-C₃ alkyl or halo;

R₄ is C₁-C₃ alkyl,

R₇ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl or 4 to 7-membered heterocycloalkyl,optionally substituted with one or more R_(s)

R₈ is C₁-C₆ alkyl;

R_(h) is -Q_(h)-T_(h), wherein Q_(h) is a bond, a C₁-C₃ alkyl linker orN(R_(N)); T_(h) is OR_(h1) or —NR_(h1)R_(h2), in which R_(h1) and R_(h2)are independently hydrogen or C₁-C₆ alkyl, or one of R_(h1) and R_(h2)is methyl and the other is a 6-membered N-containing heterocycloalkyloptionally substituted with one or two methyl, or together with the Natom to which they are attached, R_(h1) and R_(h2) form a 4 to7-membered heterocycloalkyl ring having 0 or 1 additional heteroatomsselected from oxygen and nitrogen, wherein said heterocycloalkyl ring isoptionally substituted with one or more R_(i);

R_(i) is C₁-C₃ alkyl, —NR_(N1)R_(N2) or a C₃-C₈ cycloalkyl or 5 or 6membered heterocycle each of which cycloalkyl or heterocycle isindependently optionally substituted with R_(j);

R_(N) is hydrogen, C₁-C₆ alkyl or C₃-C₈ cycloalkyl;

R_(j) is C₁-C₃ alkyl, —NR_(N1)R_(N2), or —NC(O)R_(N);

R_(N1) and R_(N2) are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈cycloalkyl, 5 or 6 membered heterocycle, each of which cycloalkyl orheterocycle is independently optionally substituted with R_(j).

For example, R₃ is hydrogen.

For example, R₃ is halogen, such as, for example, fluoro or chloro. Forexample, R₃ is fluoro.

For example R₄ is methyl, ethyl, propyl, or isopropyl. For example, R₄is methyl. For example, R₄ is isopropyl.

For example, R₇ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyran,and morpholinyl, and the like).

For example, R₇ is a 5 or 6 membered cycloalkyl or heterocycloalkyl.

For example, R₇ is a 6 membered cycloalkyl or heterocycloalkyl.

In some embodiments, R₇ is piperidinyl, tetrahydropyranyl, cyclopentyl,or cyclohexyl.

In some embodiments, R_(j) is methyl. In some embodiments, R_(j) is NH₂.

For example, R₈ is C₁, C₂ or C₃ alkyl. For example, R₈ is methyl. Forexample, R₈ is ethyl.

In some embodiments, Q_(h) is a bond. In others, Q_(h) is methylene.

In some embodiments, T_(h) is N(CH₃)₂.

In some embodiments, one of R_(h1) and R_(h2) is methyl and the other isa 6-membered N-containing heterocycloalkyl optionally substituted withone or two methyl. For example, the 6-membered N-containingheterocycloalkyl does not contain further heteroatoms in the ring. Forexample, the 6-membered N-containing heterocycloalkyl is not furthersubstituted besides the one or two methyl groups.

In some embodiments, R_(h1) and R_(h2), together with the N to whichthey are attached form a 6 membered ring. For example, T_(h) is selectedfrom piperidine, morpholine, piperazine, and N-methyl piperazine.

For example, T_(h) is morpholine.

In some embodiments, R_(i) is methyl or N(CH₃)₂. In some embodiments,R_(i) is C₃-C₈ cycloalkyl or 5 or 6 membered heterocycle. For example,R_(i) is a 6 membered cycloalkyl or heterocycle, substituted with zeroor one R_(j).

In some embodiments, R_(N) is H or methyl.

In certain compounds of Formula (III), compounds of formula IIIa, R₃ ishydrogen, R₄ is CH₃ and Q_(h) is methylene.

In certain compounds of formula III, compounds of formula IIIb, R₃ isfluoro, R₄ is isopropyl and Q_(h) is a bond.

In certain compounds of formula III, compounds of formula IIIc, R₃ ishydrogen, R₄ is propyl or isopropyl and Q_(h) is methylene.

In certain compounds of formula III, compounds of formula IIId, R₃ ishydrogen, R₄ is propyl or isopropyl and Q_(h) is a bond.

In certain compounds of formula III, compounds of Formula (IIIe),

wherein

R₃ is H or F

R₄ is methyl, i-propyl, or n-propyl,

R_(h) is

in which R_(i) is H, methyl, or

Representative compounds of the present invention include compoundslisted in Table 1. In the table below, each occurrence of

should be construed as

TABLE 1 Compound MS Number Structure (M + 1)⁺  1

501.39  2

543.22  3

486.21  4

529.30  5

471.30  6

474.30  7

448.25  8

563    9

464.3   10

462.4   11

558.45  12

559.35  13

517.3   14

557.4   15

561.35  16

515.4   17

544.35  18

547.35  19

448.25  20

614.4   21

614.4   22

519.4   23

519.3   24

559.35  25

562.4   26

463.3   27

516.35  28

560.3   29

491.25  30

518.25  31

558.35  32

516.35  35

502.3   36

557.35  37

618.35  38

618.35  39

572.35  40

572.35  41

517.25  42

572.4   43

572.6  44

573.40  45

477.35  46

477.30  47

530.35  48

576.40  49

573.45  50

573.40  51

576.45  52

531.25  53

531.30  54

615.55  55

573.40  56

546.40  57

615.40  58

 59

587.40  60

601.30  61

599.35  62

601.35  63

613.35  64

574.25  65

531.30  66

586.40  67

585.25  68

585.35  69

557.25  70

573.40  71

573.40  72

575.35  73

572.10  74

575.35  75

571.25  76

587.40  77

587.45  78

587.20  79

589.35  80

589.30  81

607.35  82

543.40  83

559.80  84

561.25  85

 86

585.37  87

600.30  88

587.40  89

503.40  90

517.30  91

531.35  92

545.40  93

557.35  94

559.20  95

599.35 (M + Na)  96

577.25  97

571.40  98

547.35  99

561.30 100

591.25 101

546.35 102

560.20 103

567.30 104

585.25 105

585.40 106

107

108

530.35 109

578.20 110

532.30 111

587.40 112

488.20 113

504.15 114

573.25 115

642.45 116

545.15 117

489.20 118

589.35 119

609.35 120

591.45 121

591.30 122

587.55 123

587.35 124

650.85 125

614.75 126

572.35 127

656.65 128

586.45 129

628.35 130

591.2  131

587.35 132

589.25 133

605.25 134

587.4  135

621.40 136

621.45 137

589.35 138

627.5  139

294.3  (M + H)/2 140

598.20 141

614.65 142

603.45 143

578.35 144

609.15 145

519.40 146

641.50 147

515.45 148

529.40 149

583.45 150

593.45 151

517.60 152

505.55 153

566.70 154

532.65 155

516.60 156

521.55 158

530.55 159

534.60 160

533.80 161

519.45 162

516.50 163

583.40 164

531.65 165

533.80 166

522.50 167

521.55 168

522.60 169

519.65 170

614.75 171

573.75 172

600.75 173

559.55 174

517.50 175

531.50 176

601.55 177

653.65 178

593.60 179

591.2  180

519.55 181

598.60 182

617.70 183

601.65 184

587.55 185

586.36 186

601.55 187

656.41 188

683.45 189

684.45 190

601.36 191

602.60 192

602.00 193

629.70 194

630.00 195

605.6  196

619.7  197

620.6  198

199

200

As used herein, “alkyl”, “C₁, C₂, C₃, C₄, C₅ or C₆ alkyl” or “C₁-C₆alkyl” is intended to include C₁, C₂, C₃, C₄, C₅ or C₆ straight chain(linear) saturated aliphatic hydrocarbon groups and C₃, C₄, C₅ or C₆branched saturated aliphatic hydrocarbon groups. For example, C₁-C₆alkyl is intended to include C₁, C₂, C₃, C₄, C₅ and C₆ alkyl groups.Examples of alkyl include, moieties having from one to six carbon atoms,such as, but not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl,s-butyl, t-butyl, n-pentyl, s-pentyl or n-hexyl.

In certain embodiments, a straight chain or branched alkyl has six orfewer carbon atoms (e.g., C₁-C₆ for straight chain, C₃-C₆ for branchedchain), and in another embodiment, a straight chain or branched alkylhas four or fewer carbon atoms.

As used herein, the term “cycloalkyl” refers to a saturated orunsaturated nonaromatic hydrocarbon mono- or multi-ring (e.g., fused,bridged, or spiro rings) system having 3 to 30 carbon atoms (e.g.,C₃-C₁₀). Examples of cycloalkyl include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantyl.The term “heterocycloalkyl” refers to a saturated or unsaturatednonaromatic 3-8 membered monocyclic, 7-12 membered bicyclic (fused,bridged, or spiro rings), or 11-14 membered tricyclic ring system(fused, bridged, or spiro rings) having one or more heteroatoms (such asO, N, S, or Se), unless specified otherwise. Examples ofheterocycloalkyl groups include, but are not limited to, piperidinyl,piperazinyl, pyrrolidinyl, dioxanyl, tetrahydrofuranyl, isoindolinyl,indolinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, oxiranyl, azetidinyl, oxetanyl,thietanyl, 1,2,3,6-tetrahydropyridinyl, tetrahydropyranyl,dihydropyranyl, pyranyl, morpholinyl, 1,4-diazepanyl, 1,4-oxazepanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl,1,4-dioxa-8-azaspiro[4.5]decanyl and the like.

The term “optionally substituted alkyl” refers to unsubstituted alkyl oralkyl having designated substituents replacing one or more hydrogenatoms on one or more carbons of the hydrocarbon backbone. Suchsubstituents can include, for example, alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, amino(including alkylamino, dialkylamino, arylamino, diarylamino andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

An “arylalkyl” or an “aralkyl” moiety is an alkyl substituted with anaryl (e.g., phenylmethyl (benzyl)). An “alkylaryl” moiety is an arylsubstituted with an alkyl (e.g., methylphenyl).

As used herein, “alkyl linker” is intended to include C₁, C₂, C₃, C₄, C₅or C₆ straight chain (linear) saturated divalent aliphatic hydrocarbongroups and C₃, C₄, C₅ or C₆ branched saturated aliphatic hydrocarbongroups. For example, C₁-C₆ alkyl linker is intended to include C₁, C₂,C₃, C₄, C₅ and C₆ alkyl linker groups. Examples of alkyl linker include,moieties having from one to six carbon atoms, such as, but not limitedto, methyl (—CH₂—), ethyl (—CH₂CH₂—), n-propyl (—CH₂CH₂CH₂—), i-propyl(—CHCH₃CH₂—), n-butyl (—CH₂CH₂CH₂CH₂—), s-butyl (—CHCH₃CH₂CH₂—), i-butyl(—C(CH₃)₂CH₂—), n-pentyl (—CH₂CH₂CH₂CH₂CH₂—), s-pentyl(—CHCH₃CH₂CH₂CH₂—) or n-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₂—).

“Alkenyl” includes unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but that contain atleast one double bond. For example, the term “alkenyl” includes straightchain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl,hexenyl, heptenyl, octenyl, nonenyl, decenyl), and branched alkenylgroups. In certain embodiments, a straight chain or branched alkenylgroup has six or fewer carbon atoms in its backbone (e.g., C₂-C₆ forstraight chain, C₃-C₆ for branched chain). The term “C₂-C₆” includesalkenyl groups containing two to six carbon atoms. The term “C₃-C₆”includes alkenyl groups containing three to six carbon atoms.

The term “optionally substituted alkenyl” refers to unsubstitutedalkenyl or alkenyl having designated substituents replacing one or morehydrogen atoms on one or more hydrocarbon backbone carbon atoms. Suchsubstituents can include, for example, alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, amino(including alkylamino, dialkylamino, arylamino, diarylamino andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

“Alkynyl” includes unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but which containat least one triple bond. For example, “alkynyl” includes straight chainalkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl,heptynyl, octynyl, nonynyl, decynyl), and branched alkynyl groups. Incertain embodiments, a straight chain or branched alkynyl group has sixor fewer carbon atoms in its backbone (e.g., C₂-C₆ for straight chain,C₃-C₆ for branched chain). The term “C₂-C₆” includes alkynyl groupscontaining two to six carbon atoms. The term “C₃-C₆” includes alkynylgroups containing three to six carbon atoms.

The term “optionally substituted alkynyl” refers to unsubstitutedalkynyl or alkynyl having designated substituents replacing one or morehydrogen atoms on one or more hydrocarbon backbone carbon atoms. Suchsubstituents can include, for example, alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, amino(including alkylamino, dialkylamino, arylamino, diarylamino andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

Other optionally substituted moieties (such as optionally substitutedcycloalkyl, heterocycloalkyl, aryl, or heteroaryl) include both theunsubstituted moieties and the moieties having one or more of thedesignated substituents. For example, substituted heterocycloalkylincludes those substituted with one or more alkyl groups, such as2,2,6,6-tetramethyl-piperidinyl and2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl.

“Aryl” includes groups with aromaticity, including “conjugated,” ormulticyclic systems with at least one aromatic ring and do not containany heteroatom in the ring structure. Examples include phenyl, benzyl,1,2,3,4-tetrahydronaphthalenyl, etc.

“Heteroaryl” groups are aryl groups, as defined above, except havingfrom one to four heteroatoms in the ring structure, and may also bereferred to as “aryl heterocycles” or “heteroaromatics.” As used herein,the term “heteroaryl” is intended to include a stable 5-, 6-, or7-membered monocyclic or 7-, 8-, 9-, 10-, 11- or 12-membered bicyclicaromatic heterocyclic ring which consists of carbon atoms and one ormore heteroatoms, e.g., 1 or 1-2 or 1-3 or 1-4 or 1-5 or 1-6heteroatoms, or e.g. 1, 2, 3, 4, 5, or 6 heteroatoms, independentlyselected from the group consisting of nitrogen, oxygen and sulfur. Thenitrogen atom may be substituted or unsubstituted (i.e., N or NR whereinR is H or other substituents, as defined). The nitrogen and sulfurheteroatoms may optionally be oxidized (i.e., N—O and S(O)_(p), wherep=1 or 2). It is to be noted that total number of S and O atoms in thearomatic heterocycle is not more than 1.

Examples of heteroaryl groups include pyrrole, furan, thiophene,thiazole, isothiazole, imidazole, triazole, tetrazole, pyrazole,oxazole, isoxazole, pyridine, pyrazine, pyridazine, pyrimidine, and thelike.

Furthermore, the terms “aryl” and “heteroaryl” include multicyclic aryland heteroaryl groups, e.g., tricyclic, bicyclic, e.g., naphthalene,benzoxazole, benzodioxazole, benzothiazole, benzoimidazole,benzothiophene, methylenedioxyphenyl, quinoline, isoquinoline,naphthrydine, indole, benzofuran, purine, benzofuran, deazapurine,indolizine.

In the case of multicyclic aromatic rings, only one of the rings needsto be aromatic (e.g., 2,3-dihydroindole), although all of the rings maybe aromatic (e.g., quinoline). The second ring can also be fused orbridged.

The cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring can besubstituted at one or more ring positions (e.g., the ring-forming carbonor heteroatom such as N) with such substituents as described above, forexample, alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkoxy,alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl,aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylthiocarbonyl, phosphate, phosphonato, phosphinato, amino (includingalkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moiety. Aryl and heteroarylgroups can also be fused or bridged with alicyclic or heterocyclicrings, which are not aromatic so as to form a multicyclic system (e.g.,tetralin, methylenedioxyphenyl).

As used herein, “carbocycle” or “carbocyclic ring” is intended toinclude any stable monocyclic, bicyclic or tricyclic ring having thespecified number of carbons, any of which may be saturated, unsaturated,or aromatic. Carbocycle includes cycloalkyl and aryl. For example, aC₃-C₁₄ carbocycle is intended to include a monocyclic, bicyclic ortricyclic ring having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbonatoms. Examples of carbocycles include, but are not limited to,cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl,cyclohexyl, cycloheptenyl, cycloheptyl, cycloheptenyl, adamantyl,cyclooctyl, cyclooctenyl, cyclooctadienyl, fluorenyl, phenyl, naphthyl,indanyl, adamantyl and tetrahydronaphthyl. Bridged rings are alsoincluded in the definition of carbocycle, including, for example,[3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane and[2.2.2]bicyclooctane. A bridged ring occurs when one or more carbonatoms link two non-adjacent carbon atoms. In one embodiment, bridgerings are one or two carbon atoms. It is noted that a bridge alwaysconverts a monocyclic ring into a tricyclic ring. When a ring isbridged, the substituents recited for the ring may also be present onthe bridge. Fused (e.g., naphthyl, tetrahydronaphthyl) and spiro ringsare also included.

As used herein, “heterocycle” or “heterocyclic group” includes any ringstructure (saturated, unsaturated, or aromatic) which contains at leastone ring heteroatom (e.g., N, O or S). Heterocycle includesheterocycloalkyl and heteroaryl. Examples of heterocycles include, butare not limited to, morpholine, pyrrolidine, tetrahydrothiophene,piperidine, piperazine, oxetane, pyran, tetrahydropyran, azetidine, andtetrahydrofuran.

Examples of heterocyclic groups include, but are not limited to,acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl,benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl,benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl,chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl,indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl,isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl,isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl,naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl,1,2,4-oxadiazol5(4H)-one, oxazolidinyl, oxazolyl, oxindolyl,pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl,piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl,pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl,pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl,pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl,quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl,1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl.

The term “substituted,” as used herein, means that any one or morehydrogen atoms on the designated atom is replaced with a selection fromthe indicated groups, provided that the designated atom's normal valencyis not exceeded, and that the substitution results in a stable compound.When a substituent is oxo or keto (i.e., ═O), then 2 hydrogen atoms onthe atom are replaced. Keto substituents are not present on aromaticmoieties. Ring double bonds, as used herein, are double bonds that areformed between two adjacent ring atoms (e.g., C═C, C═N or N═N). “Stablecompound” and “stable structure” are meant to indicate a compound thatis sufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

When a bond to a substituent is shown to cross a bond connecting twoatoms in a ring, then such substituent may be bonded to any atom in thering. When a substituent is listed without indicating the atom via whichsuch substituent is bonded to the rest of the compound of a givenformula, then such substituent may be bonded via any atom in suchformula. Combinations of substituents and/or variables are permissible,but only if such combinations result in stable compounds.

When any variable (e.g., R₁) occurs more than one time in anyconstituent or formula for a compound, its definition at each occurrenceis independent of its definition at every other occurrence. Thus, forexample, if a group is shown to be substituted with 0-2 R₁ moieties,then the group may optionally be substituted with up to two R₁ moietiesand R₁ at each occurrence is selected independently from the definitionof R₁. Also, combinations of substituents and/or variables arepermissible, but only if such combinations result in stable compounds.

The term “hydroxy” or “hydroxyl” includes groups with an —OH or —O⁻.

As used herein, “halo” or “halogen” refers to fluoro, chloro, bromo andiodo. The term “perhalogenated” generally refers to a moiety wherein allhydrogen atoms are replaced by halogen atoms. The term “haloalkyl” or“haloalkoxyl” refers to an alkyl or alkoxyl substituted with one or morehalogen atoms.

The term “carbonyl” includes compounds and moieties which contain acarbon connected with a double bond to an oxygen atom. Examples ofmoieties containing a carbonyl include, but are not limited to,aldehydes, ketones, carboxylic acids, amides, esters, anhydrides, etc.

The term “carboxyl” refers to —COOH or its C₁-C₆ alkyl ester.

“Acyl” includes moieties that contain the acyl radical (R—C(O)—) or acarbonyl group. “Substituted acyl” includes acyl groups where one ormore of the hydrogen atoms are replaced by, for example, alkyl groups,alkynyl groups, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, amino (including alkylamino, dialkylamino,arylamino, diarylamino and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

“Aroyl” includes moieties with an aryl or heteroaromatic moiety bound toa carbonyl group. Examples of aroyl groups include phenylcarboxy,naphthyl carboxy, etc.

“Alkoxyalkyl,” “alkylaminoalkyl,” and “thioalkoxyalkyl” include alkylgroups, as described above, wherein oxygen, nitrogen, or sulfur atomsreplace one or more hydrocarbon backbone carbon atoms.

The term “alkoxy” or “alkoxyl” includes substituted and unsubstitutedalkyl, alkenyl and alkynyl groups covalently linked to an oxygen atom.Examples of alkoxy groups or alkoxyl radicals include, but are notlimited to, methoxy, ethoxy, isopropyloxy, propoxy, butoxy and pentoxygroups. Examples of substituted alkoxy groups include halogenated alkoxygroups. The alkoxy groups can be substituted with groups such asalkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, amino (including alkylamino, dialkylamino,arylamino, diarylamino, and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moieties. Examples of halogen substituted alkoxygroups include, but are not limited to, fluoromethoxy, difluoromethoxy,trifluoromethoxy, chloromethoxy, dichloromethoxy and trichloromethoxy.

The term “ether” or “alkoxy” includes compounds or moieties whichcontain an oxygen bonded to two carbon atoms or heteroatoms. Forexample, the term includes “alkoxyalkyl,” which refers to an alkyl,alkenyl, or alkynyl group covalently bonded to an oxygen atom which iscovalently bonded to an alkyl group.

The term “ester” includes compounds or moieties which contain a carbonor a heteroatom bound to an oxygen atom which is bonded to the carbon ofa carbonyl group. The term “ester” includes alkoxycarboxy groups such asmethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl,pentoxycarbonyl, etc.

The term “thioalkyl” includes compounds or moieties which contain analkyl group connected with a sulfur atom. The thioalkyl groups can besubstituted with groups such as alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, carboxy acid, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, amino (includingalkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moieties.

The term “thiocarbonyl” or “thiocarboxy” includes compounds and moietieswhich contain a carbon connected with a double bond to a sulfur atom.

The term “thioether” includes moieties which contain a sulfur atombonded to two carbon atoms or heteroatoms. Examples of thioethersinclude, but are not limited to alkthioalkyls, alkthioalkenyls, andalkthioalkynyls. The term “alkthioalkyls” include moieties with analkyl, alkenyl, or alkynyl group bonded to a sulfur atom which is bondedto an alkyl group. Similarly, the term “alkthioalkenyls” refers tomoieties wherein an alkyl, alkenyl or alkynyl group is bonded to asulfur atom which is covalently bonded to an alkenyl group; andalkthioalkynyls” refers to moieties wherein an alkyl, alkenyl or alkynylgroup is bonded to a sulfur atom which is covalently bonded to analkynyl group.

As used herein, “amine” or “amino” refers to unsubstituted orsubstituted —NH₂. “Alkylamino” includes groups of compounds whereinnitrogen of —NH₂ is bound to at least one alkyl group. Examples ofalkylamino groups include benzylamino, methylamino, ethylamino,phenethylamino, etc. “Dialkylamino” includes groups wherein the nitrogenof —NH₂ is bound to at least two additional alkyl groups. Examples ofdialkylamino groups include, but are not limited to, dimethylamino anddiethylamino. “Arylamino” and “diarylamino” include groups wherein thenitrogen is bound to at least one or two aryl groups, respectively.“Aminoaryl” and “aminoaryloxy” refer to aryl and aryloxy substitutedwith amino. “Alkylarylamino,” “alkylaminoaryl” or “arylaminoalkyl”refers to an amino group which is bound to at least one alkyl group andat least one aryl group. “Alkaminoalkyl” refers to an alkyl, alkenyl, oralkynyl group bound to a nitrogen atom which is also bound to an alkylgroup. “Acylamino” includes groups wherein nitrogen is bound to an acylgroup. Examples of acylamino include, but are not limited to,alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido groups.

The term “amide” or “aminocarboxy” includes compounds or moieties thatcontain a nitrogen atom that is bound to the carbon of a carbonyl or athiocarbonyl group. The term includes “alkaminocarboxy” groups thatinclude alkyl, alkenyl or alkynyl groups bound to an amino group whichis bound to the carbon of a carbonyl or thiocarbonyl group. It alsoincludes “arylaminocarboxy” groups that include aryl or heteroarylmoieties bound to an amino group that is bound to the carbon of acarbonyl or thiocarbonyl group. The terms “alkylaminocarboxy”,“alkenylaminocarboxy”, “alkynylaminocarboxy” and “arylaminocarboxy”include moieties wherein alkyl, alkenyl, alkynyl and aryl moieties,respectively, are bound to a nitrogen atom which is in turn bound to thecarbon of a carbonyl group. Amides can be substituted with substituentssuch as straight chain alkyl, branched alkyl, cycloalkyl, aryl,heteroaryl or heterocycle. Substituents on amide groups may be furthersubstituted.

Compounds of the present invention that contain nitrogens can beconverted to N-oxides by treatment with an oxidizing agent (e.g.,3-chloroperoxybenzoic acid (mCPBA) and/or hydrogen peroxides) to affordother compounds of the present invention. Thus, all shown and claimednitrogen-containing compounds are considered, when allowed by valencyand structure, to include both the compound as shown and its N-oxidederivative (which can be designated as N→O or N⁺—O⁻). Furthermore, inother instances, the nitrogens in the compounds of the present inventioncan be converted to N-hydroxy or N-alkoxy compounds. For example,N-hydroxy compounds can be prepared by oxidation of the parent amine byan oxidizing agent such as m-CPBA. All shown and claimednitrogen-containing compounds are also considered, when allowed byvalency and structure, to cover both the compound as shown and itsN-hydroxy (i.e., N—OH) and N-alkoxy (i.e., N—OR, wherein R issubstituted or unsubstituted C₁-C₆ alkyl, C₁-C₆ alkenyl, C₁-C₆ alkynyl,3-14-membered carbocycle or 3-14-membered heterocycle) derivatives.

In the present specification, the structural formula of the compoundrepresents a certain isomer for convenience in some cases, but thepresent invention includes all isomers, such as geometrical isomers,optical isomers based on an asymmetrical carbon, stereoisomers,tautomers, and the like, it being understood that not all isomers mayhave the same level of activity. In addition, a crystal polymorphism maybe present for the compounds represented by the formula. It is notedthat any crystal form, crystal form mixture, or anhydride or hydratethereof is included in the scope of the present invention. Furthermore,so-called metabolite which is produced by degradation of the presentcompound in vivo is included in the scope of the present invention.

“Isomerism” means compounds that have identical molecular formulae butdiffer in the sequence of bonding of their atoms or in the arrangementof their atoms in space. Isomers that differ in the arrangement of theiratoms in space are termed “stereoisomers.” Stereoisomers that are notmirror images of one another are termed “diastereoisomers,” andstereoisomers that are non-superimposable mirror images of each otherare termed “enantiomers” or sometimes optical isomers. A mixturecontaining equal amounts of individual enantiomeric forms of oppositechirality is termed a “racemic mixture.”

A carbon atom bonded to four nonidentical substituents is termed a“chiral center.”

“Chiral isomer” means a compound with at least one chiral center.Compounds with more than one chiral center may exist either as anindividual diastereomer or as a mixture of diastereomers, termed“diastereomeric mixture.” When one chiral center is present, astereoisomer may be characterized by the absolute configuration (R or S)of that chiral center. Absolute configuration refers to the arrangementin space of the substituents attached to the chiral center. Thesubstituents attached to the chiral center under consideration areranked in accordance with the Sequence Rule of Cahn, Ingold and Prelog.(Cahn et al., Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahnet al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J. Chem. Soc. 1951(London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn, J. Chem.Educ. 1964, 41, 116).

“Geometric isomer” means the diastereomers that owe their existence tohindered rotation about double bonds or a cycloalkyl linker (e.g.,1,3-cylcobutyl). These configurations are differentiated in their namesby the prefixes cis and trans, or Z and E, which indicate that thegroups are on the same or opposite side of the double bond in themolecule according to the Cahn-Ingold-Prelog rules.

It is to be understood that the compounds of the present invention maybe depicted as different chiral isomers or geometric isomers. It shouldalso be understood that when compounds have chiral isomeric or geometricisomeric forms, all isomeric forms are intended to be included in thescope of the present invention, and the naming of the compounds does notexclude any isomeric forms, it being understood that not all isomers mayhave the same level of activity.

Furthermore, the structures and other compounds discussed in thisinvention include all atropic isomers thereof, it being understood thatnot all atropic isomers may have the same level of activity. “Atropicisomers” are a type of stereoisomer in which the atoms of two isomersare arranged differently in space. Atropic isomers owe their existenceto a restricted rotation caused by hindrance of rotation of large groupsabout a central bond. Such atropic isomers typically exist as a mixture,however as a result of recent advances in chromatography techniques, ithas been possible to separate mixtures of two atropic isomers in selectcases.

“Tautomer” is one of two or more structural isomers that exist inequilibrium and is readily converted from one isomeric form to another.This conversion results in the formal migration of a hydrogen atomaccompanied by a switch of adjacent conjugated double bonds. Tautomersexist as a mixture of a tautomeric set in solution. In solutions wheretautomerization is possible, a chemical equilibrium of the tautomerswill be reached. The exact ratio of the tautomers depends on severalfactors, including temperature, solvent and pH. The concept of tautomersthat are interconvertable by tautomerizations is called tautomerism.

Of the various types of tautomerism that are possible, two are commonlyobserved. In keto-enol tautomerism a simultaneous shift of electrons anda hydrogen atom occurs. Ring-chain tautomerism arises as a result of thealdehyde group (—CHO) in a sugar chain molecule reacting with one of thehydroxy groups (—OH) in the same molecule to give it a cyclic(ring-shaped) form as exhibited by glucose.

Common tautomeric pairs are: ketone-enol, amide-nitrile, lactam-lactim,amide-imidic acid tautomerism in heterocyclic rings (e.g., innucleobases such as guanine, thymine and cytosine), imine-enamine andenamine-enamine. An example of keto-enol equilibria is betweenpyridin-2(1H)-ones and the corresponding pyridin-2-ols, as shown below.

It is to be understood that the compounds of the present invention maybe depicted as different tautomers. It should also be understood thatwhen compounds have tautomeric forms, all tautomeric forms are intendedto be included in the scope of the present invention, and the naming ofthe compounds does not exclude any tautomer form. It will be understoodthat certain tautomers may have a higher level of activity than others.

The term “crystal polymorphs”, “polymorphs” or “crystal forms” meanscrystal structures in which a compound (or a salt or solvate thereof)can crystallize in different crystal packing arrangements, all of whichhave the same elemental composition. Different crystal forms usuallyhave different X-ray diffraction patterns, infrared spectral, meltingpoints, density hardness, crystal shape, optical and electricalproperties, stability and solubility. Recrystallization solvent, rate ofcrystallization, storage temperature, and other factors may cause onecrystal form to dominate. Crystal polymorphs of the compounds can beprepared by crystallization under different conditions.

The compounds of any of Formulae disclosed herein include the compoundsthemselves, as well as their salts, their esters, their solvates, andtheir prodrugs, if applicable. A salt, for example, can be formedbetween an anion and a positively charged group (e.g., amino) on anaryl- or heteroaryl-substituted benzene compound. Suitable anionsinclude chloride, bromide, iodide, sulfate, bisulfate, sulfamate,nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate,glutamate, glucuronate, glutarate, malate, maleate, succinate, fumarate,tartrate, tosylate, salicylate, lactate, naphthalenesulfonate, andacetate (e.g., trifluoroacetate). The term “pharmaceutically acceptableanion” refers to an anion suitable for forming a pharmaceuticallyacceptable salt. Likewise, a salt can also be formed between a cationand a negatively charged group (e.g., carboxylate) on an aryl- orheteroaryl-substituted benzene compound. Suitable cations include sodiumion, potassium ion, magnesium ion, calcium ion, and an ammonium cationsuch as tetramethylammonium ion. The aryl- or heteroaryl-substitutedbenzene compounds also include those salts containing quaternarynitrogen atoms. Examples of prodrugs include esters and otherpharmaceutically acceptable derivatives, which, upon administration to asubject, are capable of providing active aryl- or heteroaryl-substitutedbenzene compounds.

Additionally, the compounds of the present invention, for example, thesalts of the compounds, can exist in either hydrated or unhydrated (theanhydrous) form or as solvates with other solvent molecules. Nonlimitingexamples of hydrates include monohydrates, dihydrates, etc. Nonlimitingexamples of solvates include ethanol solvates, acetone solvates, etc.

“Solvate” means solvent addition forms that contain eitherstoichiometric or non stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate; and if the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one molecule of the substance inwhich the water retains its molecular state as H₂O.

As used herein, the term “analog” refers to a chemical compound that isstructurally similar to another but differs slightly in composition (asin the replacement of one atom by an atom of a different element or inthe presence of a particular functional group, or the replacement of onefunctional group by another functional group). Thus, an analog is acompound that is similar or comparable in function and appearance, butnot in structure or origin to the reference compound.

As defined herein, the term “derivative” refers to compounds that have acommon core structure, and are substituted with various groups asdescribed herein. For example, all of the compounds represented byFormula (I) are aryl- or heteroaryl-substituted benzene compounds, andhave Formula (I) as a common core.

The term “bioisostere” refers to a compound resulting from the exchangeof an atom or of a group of atoms with another, broadly similar, atom orgroup of atoms. The objective of a bioisosteric replacement is to createa new compound with similar biological properties to the parentcompound. The bioisosteric replacement may be physicochemically ortopologically based. Examples of carboxylic acid bioisosteres include,but are not limited to, acyl sulfonimides, tetrazoles, sulfonates andphosphonates. See, e.g., Patani and LaVoie, Chem. Rev. 96, 3147-3176,1996.

The present invention is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include tritium anddeuterium, and isotopes of carbon include C-13 and C-14.

2. SYNTHESIS OF ARYL- OR HETEROARYL-SUBSTITUTED BENZENE COMPOUNDS

The present invention provides methods for the synthesis of thecompounds of any Formula disclosed herein. The present invention alsoprovides detailed methods for the synthesis of various disclosedcompounds of the present invention according to the following schemes asshown in the Examples.

Throughout the description, where compositions are described as having,including, or comprising specific components, it is contemplated thatcompositions also consist essentially of, or consist of, the recitedcomponents. Similarly, where methods or processes are described ashaving, including, or comprising specific process steps, the processesalso consist essentially of, or consist of, the recited processingsteps. Further, it should be understood that the order of steps or orderfor performing certain actions is immaterial so long as the inventionremains operable. Moreover, two or more steps or actions can beconducted simultaneously.

The synthetic processes of the invention can tolerate a wide variety offunctional groups, therefore various substituted starting materials canbe used. The processes generally provide the desired final compound ator near the end of the overall process, although it may be desirable incertain instances to further convert the compound to a pharmaceuticallyacceptable salt, ester, or prodrug thereof.

Compounds of the present invention can be prepared in a variety of waysusing commercially available starting materials, compounds known in theliterature, or from readily prepared intermediates, by employingstandard synthetic methods and procedures either known to those skilledin the art, or which will be apparent to the skilled artisan in light ofthe teachings herein. Standard synthetic methods and procedures for thepreparation of organic molecules and functional group transformationsand manipulations can be obtained from the relevant scientificliterature or from standard textbooks in the field. Although not limitedto any one or several sources, classic texts such as Smith, M. B.,March, J., March's Advanced Organic Chemistry: Reactions, Mechanisms,and Structure, 5^(th) edition, John Wiley & Sons: New York, 2001;Greene, T. W., Wuts, P. G. M., Protective Groups in Organic Synthesis,3^(rd) edition, John Wiley & Sons: New York, 1999; R. Larock,Comprehensive Organic Transformations, VCH Publishers (1989); L. Fieserand M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, JohnWiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagentsfor Organic Synthesis, John Wiley and Sons (1995), incorporated byreference herein, are useful and recognized reference textbooks oforganic synthesis known to those in the art. The following descriptionsof synthetic methods are designed to illustrate, but not to limit,general procedures for the preparation of compounds of the presentinvention.

Compounds of the present invention can be conveniently prepared by avariety of methods familiar to those skilled in the art. The compoundsof this invention with any Formula disclosed herein may be preparedaccording to the procedures illustrated in Schemes 1-10 below, fromcommercially available starting materials or starting materials whichcan be prepared using literature procedures. The Z and R groups (such asR₂, R₃, R₄, R₆, R₇, R₈, and R₁₂) in Schemes 1-10 are as defined in anyof Formulae disclosed herein, unless otherwise specified.

One of ordinary skill in the art will note that, during the reactionsequences and synthetic schemes described herein, the order of certainsteps may be changed, such as the introduction and removal of protectinggroups.

One of ordinary skill in the art will recognize that certain groups mayrequire protection from the reaction conditions via the use ofprotecting groups. Protecting groups may also be used to differentiatesimilar functional groups in molecules. A list of protecting groups andhow to introduce and remove these groups can be found in Greene, T. W.,Wuts, P. G. M., Protective Groups in Organic Synthesis, 3^(rd) edition,John Wiley & Sons: New York, 1999.

Preferred protecting groups include, but are not limited to:

For a hydroxyl moiety: TBS, benzyl, THP, Ac

For carboxylic acids: benzyl ester, methyl ester, ethyl ester, allylester

For amines: Cbz, BOC, DMB

For diols: Ac (×2) TBS (×2), or when taken together acetonides

For thiols: Ac

For benzimidazoles: SEM, benzyl, PMB, DMB

For aldehydes: di-alkyl acetals such as dimethoxy acetal or diethylacetyl.

In the reaction schemes described herein, multiple stereoisomers may beproduced. When no particular stereoisomer is indicated, it is understoodto mean all possible stereoisomers that could be produced from thereaction. A person of ordinary skill in the art will recognize that thereactions can be optimized to give one isomer preferentially, or newschemes may be devised to produce a single isomer. If mixtures areproduced, techniques such as preparative thin layer chromatography,preparative HPLC, preparative chiral HPLC, or preparative SFC may beused to separate the isomers.

The following abbreviations are used throughout the specification andare defined below:

-   -   AA ammonium acetate    -   ACN acetonitrile    -   Ac acetyl    -   AcOH acetic acid    -   atm atmosphere    -   aq. aqueous    -   BID or b.i.d. bis in die (twice a day)    -   tBuOK potassium t-butoxide    -   Bn benzyl    -   BOC tert-butoxy carbonyl    -   BOP        (benzotriazol-1-yloxy)tris(dimethylamino)-phosphoniumhexafluorophosphate    -   Cbz benzyloxy carbonyl    -   CDCl₃ deuterated chloroform    -   CH₂Cl₂ dichloromethane    -   COMU        (1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethyl-amino-morpholino-carbenium        hexafluorophosphate    -   d days    -   DBU 1,8-diazabicyclo[5.4.0]undec-7-ene    -   DCE 1,2 dichloroethane    -   DCM dichloromethane    -   DEAD Diethyl azodicarboxylate    -   DIAD Diisopropyl azodicarboxylate    -   DiBAL-H diisobutyl aluminium hydride    -   DIPEA N,N-diisopropylethylamine (Hunig's base)    -   DMA Dimethylacetamide    -   DMAP N, N dimethyl-4-aminopyridine    -   DMB 2,4 dimethoxy benzyl    -   DMF N,NDimethylformamide    -   DMSO Dimethyl sulfoxide    -   DPPA Diphenylphosphonic azide    -   EA or EtOAc Ethyl acetate    -   EDC or EDCI N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide    -   Et₂O diethyl ether    -   ELS Evaporative Light Scattering    -   ESI− Electrospray negative mode    -   ESI+ Electrospray positive mode    -   Et₃N or TEA triethylamine    -   EtOH ethanol    -   FA formic acid    -   FC or FCC Flash chromatography    -   h hours    -   H₂O water    -   HATU O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluorophosphate    -   HOAT 1-Hydroxy-7-azabenzotriazole    -   HOBt 1-Hydroxybenzotriazole    -   HO-Su N-Hydroxysuccinimide    -   HCl hydrogen chloride or hydrochloric acid    -   HPLC High performance liquid chromatography    -   K₂CO₃ potassium carbonate    -   KHMDs Potassium hexamethyldisilazide    -   LC/MS or LC-MS Liquid chromatography mass spectrum    -   LDA Lithium diisopropylamide    -   LiHMDs Lithium hexamethyldisilazide    -   LG leaving group    -   M Molar    -   m/z mass/charge ratio    -   m-CPBA meta-chloroperbenzoic acid    -   MeCN Acetonitrile    -   MeOD d4-methanol    -   MeI Methyl iodide    -   MS3 Å 3 Å molecular sieves    -   MgSO₄ Magnesium Sulfate    -   min minutes    -   Ms Mesyl    -   MsCl Mesyl chloride    -   MsO Mesylate    -   MS Mass Spectrum    -   MWI microwave irradiation    -   Na₂CO₃ sodium carbonate    -   Na₂SO₄ sodium sulfate    -   NaHCO₃ sodium bicarbonate    -   NaHMDs Sodium hexamethyldisilazide    -   NaOH sodium hydroxide    -   NaHCO₃ sodium bicarbonate    -   Na₂SO₄ sodium sulfate    -   NIS N-iodosuccinimide    -   NMR Nuclear Magnetic Resonance    -   o/n or O/N overnight    -   Pd/C Palladium on carbon    -   Pd(dppf)Cl₂.DCM        [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II),        complex with dichloromethane    -   PPAA 1-Propanephosphonic acid cyclic anhydride    -   Pd(OH)₂ Palladium dihydroxide    -   PE Petroleum Ether    -   PG protecting group    -   PMB para methoxybenzyl    -   p.o. per os (oral administration)    -   ppm parts per million    -   prep HPLC preparative High Performance Liquid Chromatography    -   prep TLC preparative thin layer chromatography    -   p-TsOH para-toluenesulfonic acid    -   PyBOP (Benzotriazol-1-yloxy)tripyrrolidinophosphonium        hexafluorophosphate    -   QD or q.d. quaque die (once a day)    -   RBF round bottom flask    -   RP-HPLC Reverse phase High Performance liquid chromatography    -   Rt or RT Room temperature    -   SEM (Trimethylsilyl)ethoxymethyl    -   SEMCl (Trimethylsilyl)ethoxymethyl chloride    -   SFC Super critical chromatography    -   SGC silica gel chromatography    -   STAB Sodium triacetoxy borohydride    -   TBAF tetra-n-butylammonium fluoride    -   TBME tert-Butyl methyl ether    -   TEA Triethylamine    -   TFA trifluoroacetic acid    -   TfO triflate    -   THF tetrahydrofuran    -   THP tetrahydropyran    -   TID or t.i.d ter in die (three times a day)    -   TLC thin layer chromatography    -   TMSCl Trimethylsilyl chloride    -   Ts tosyl    -   TsOH tosic acid    -   UV ultraviolet

Scheme 1 shows the synthesis of modified aryl analogs following ageneral route that utilizes well-established chemistry. Substitutednitrobenzoic acids, many of which are commercially available or can bemade nitrations of the appropriate substituted benzoic acids or otherchemistry known to ones skilled in the art, can be converted to theirmethyl esters by treatment with methyliodide in a polar solvent such asDMF in the presence of an appropriate base such as sodium carbonate atan appropriate temperature such as 60° C. (Step 1). The nitro group canbe reduced to an amine using an appropriate reducing agent such as ironin the presence of an acid such as ammonium chloride in a protic solventsuch as ethanol at an appropriate temperature such as 80° C. (Step 2).Introduction of the R₇ can be done using a reductive amination with anappropriate ketone or aldehyde in the presence of an appropriatereducing agent such as sodium cyanoborohydride and catalytic acid suchas acetic acid in an appropriate solvent such as methanol. A variety ofR₈ groups can be introduced by alkylation using R₈-LG, where LG is aleaving group such as iodine, in the presence of a mild base such ascesium carbonate in an appropriate polar solvent such as acetonitrile atan appropriate temperature such as 80° C. (Step 4). Alternatively, R₈groups can be introduced by reductive amination with R₈-ketone orR₈-aldehyde in the presence of an appropriate reducing agent such assodium cyanoborohydride and catalytic acid such as acetic acid in anappropriate solvent such as methanol. The ester moiety can be convertedto an amide using a standard two step protocol. The ester can behydrolyzed to the corresponding acid using a suitable base such assodium hydroxide in a polar solvent such as ethanol (Step 5). The acidwould then be subjecting to a standard amide coupling reaction whereuponthe appropriate amine would be added along with a suitable amidecoupling reagent such as PYBOP in a suitable solvent such as DMSO togive the desired amide (Step 6).

Depending upon the nature of the R₆ substituent, further chemicalmodification could be employed to convert the R₆ substituent into analternative R₆ substituent. A representative sampling of suchmodifications could include hydrogenation, protecting group removalfollowed by additional amide coupling reactions, palladium catalyzedcoupling reactions, reductive amination reactions or alkylationreactions. For example, as depicted in Scheme 2, if R₆ is a bromide,alternative R₆ substituents could then be introduced using standardtransition metal-based protocols that rely upon a leaving group such asa bromide as a connection point. The bromide would be combined with anappropriate boronic ester derivative, in the presence of a mild base anda palladium catalyst in a polar solvent such as dioxane/water, atelevated temperature to give the desired new R₆ substituent (i.e. Suzukireaction). For example, as depicted in Scheme 3, if the Suzuki reactionis conducted with a boronic ester derivative bearing a formyl groupfurther modification by reductive amination reaction with primary andsecondary amines (e.g. morpholine, dimethylamine) can be conducted tointroduce amine groups.

Depending upon the nature of the R₇ substituent, further chemicalmodification subsequent to Step 6 of Scheme 1 could be employed toconvert the R₇ substituent into an alternative R₇ substituent. Forexample a protected amino group contained within R₇ may be subjected todeprotection reaction (e.g. Boc group cleavage) to give free aminogroups. Such free amino groups may be subjected to reductive aminationreactions or alkylation reactions to give substituted amines.

Scheme 4 shows the general synthesis of 2,6-disubstitutedisonicotinamide compounds. Suzuki reaction in Step 1 of an aryl boronicacid compound with methyl 2,6-dichloroisonicotinate starting materialcan be used to introduce an aryl group which may be substituted with afunctional group X that is suitable for further transformation. Such Xgroups include formyl or hydroxymethyl which can readily be transformedin Step 2 to various groups Y. Such Y groups include aminomethyl,monoalkylaminomethyl and dialkylaminomethyl groups. The latter can beprepared by reductive amination in the case where X is formyl or byconverting X=hydroxymethyl to bromomethyl followed by alkylation with anamine. Ester hydrolysis a subsequent step gives an acid intermediatewhich can be coupled with appropriate 3-(aminomethyl)-pyridin-2(1H)-onesto give the penultimate 2-chloro-6-aryl-isonicotine amide intermediate.Suzuki reaction or amination reaction then gives compounds substitutedin the 2-position with a Z group. In the case of an amination reactionexamples of Z can be monoalkylamino or dialkylamino. In the case of aSuzuki reaction Z can be aryl, dihydroaryl or tetrahydroaryl such ascyclohexenyl.

Scheme 5 shows the general synthesis of 6-aryl-3-methyl-picolinamideshaving monoalkylamino or dialkylamino groups in the 4-position. Startingfrom methyl 3-bromo-6-chloropicolinate oxidation to the N-oxide followedby chlorination with phosphorus oxychloride gives methyl3-bromo-4,6-dichloropicolinate. The 4-chloro group can be selectivelysubstituted with diverse mono and dialkyl amines which may also containfunctional or protected functional groups that may be unmasked at alater stage. Palladium catalyzed methylation with tetramethyltinfollowed by ester hydrolysis and amide coupling with appropriate3-(aminomethyl)-pyridin-2(1H)-ones yields penultimate 2-chloro pyridineintermediates. Suzuki coupling reaction group of these intermediateswith aryl boronic acids results in replacement of the 2-chloro groupwith an aryl group. Thus, this yields 6-aryl-3-methyl-picolinamideshaving monoalkylamino or dialkylamino groups in the 4-position. The arylgroup which may be substituted with a functional group X that remains inthe final product or is converted to an another group by deprotection orfunctional group conversion reaction e.g. reductive amination.

General syntheses of 3-(aminomethyl)-pyridin-2(1H)-ones intermediatesfor the amide coupling reaction from Scheme 1 are depicted in Scheme 6below. In one method, a diketone can be condensed with 2-cyanoacetamidein the presence of an appropriate reagent such as piperidine acetate ina polar solvent such as ethanol to provide a cyanopyridone (Step 9). Inanother method, when R₃ is H, an appropriately substituted alkynylketone can be condensed with 2-cyanoacetamide in the presence of anappropriate reagent such as piperidine acetate in a polar solvent suchas ethanol to provide a cyanopyridone (Step 11). The cyano group can bereduced under appropriate conditions such as hydrogenation in thepresence of catalytic Raney nickel in a polar solvent such as ammoniumin methanol to provide the amine (Step 10).

Additionally, depending upon the nature of the R₂, R₃, or R₄ group,further chemical modification can be employed to convert each of themindependently into an alternative substituent. A representative samplingof such modifications can include hydrogenation, protecting groupremoval followed by additional amide coupling reactions, palladiumcatalyzed coupling reactions, reductive amination reactions, andalkylation reactions. Scheme 4 depicts a variant of the generalsynthesis route of Scheme 1 based on 2-substituted (substituent is anR₁₂ group) methyl 3-amino-5-bromo-benzoate starting materials. Thesestarting materials can in turn be prepared from 2-substituted3-nitro-benzoic acids which are commercially available or can beprepared by nitration of 2-substituted benzoic acids. Thus, brominationof 2-substituted 3-nitro-benzoic acids with a suitable reagent such as1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione yields the appropriate2-substituted 3-nitro-5-bromo-benzoic acids. A variety of esterificationand then nitro group reduction methods can then be sequentiallyimplemented to prepare the 2-substituted methyl 3-amino-5-bromo-benzoatestarting materials from the 2-substituted 3-nitro-5-bromo-benzoic acids.

As depicted in Scheme 7 the R₇ group can be introduced from2-substituted methyl 3-amino-5-bromo-benzoates in Step 1 using areductive amination with an appropriate R₇-ketone or R₇-aldehyde in thepresence of an appropriate reducing agent such as sodiumcyanoborohydride and catalytic acid such as acetic acid in anappropriate solvent such as methanol. Similarly, R₈ groups can beintroduced in Step 2 by reductive amination with R₈-ketone orR₈-aldehyde in the presence of an appropriate reducing agent such assodium cyanoborohydride and catalytic acid such as acetic acid in anappropriate solvent such as methanol. Alternatively, a variety of R₈groups can be introduced by alkylation using R₈-LG, where LG is aleaving group such as iodine, in the presence of a mild base such ascesium carbonate in an appropriate polar solvent such as acetonitrile atan appropriate temperature such as 80° C. In Step 3, aryl groupscorresponding to R₆ can be introduced by Suzuki reaction of theintermediate bromide with an appropriate aryl boronic acid or esterderivative, e.g., X—Ar—B(OH)₂, in the presence of a mild base and apalladium catalyst in a polar solvent such as dioxane/water, at elevatedtemperature. The X group in X—Ar—B(OH)₂ may be a fully elaboratedsubstituent on the aryl ring or may be a functional group that can beconverted into another group by functional group modification. Arepresentative sampling of such modifications could includehydrogenation, protecting group removal followed by additional amidecoupling reactions, palladium catalyzed coupling reactions, reductiveamination reactions or alkylation reactions. For example if the Suzukireaction is conducted with a boronic acid derivative bearing a formylgroup further modification by reductive amination reaction with primaryand secondary amines (e.g. morpholine, dimethylamine) can be conductedto introduce amine groups. In Step 4 the ester moiety can be hydrolyzedto the corresponding acid using a suitable base such as sodium hydroxidein a polar solvent such as ethanol. In Step 5, the acid can be subjectedto a standard amide coupling reaction whereupon the appropriate aminewould be added along with a suitable amide coupling reagent such asPYBOP in a suitable solvent such as DMSO to give the desired amide.Depending upon the nature of the R₇ substituent, further chemicalmodification subsequent to Step 5 of Scheme 4 could be employed toconvert the R₇ substituent into an alternative R₇ substituent. Forexample a protected amino group contained within R₇ may be subjected todeprotection reaction (e.g. Boc group cleavage) to give free aminogroups. Such free amino groups may be subjected to reductive aminationreactions or alkylation reactions to give substituted amines.

Scheme 8 below depicts the general synthesis of 2-monoalkylamino and2-dialkylmino-3-substituted-6-aryl-isonicotinamides wherein the3-substituent corresponds to R₁₂ and the 6-aryl group corresponds to R₆,Formula I In Step 1 the 3-substituent may be introduced by the methoddescribed by Epsztain J. et al. Tetrahedron, 1991, v. 47, 1697-16708, bymetallation of 2-chloro-isonicotinanilide with n-butyllithium followedby trapping with an an alkyliodide such as methyliodide or aldehyde orother electrophilic group.

In cases where the trapping reagent yields a substituent with afunctional group this group may be masked or converted into anotherfunctional group compatible with the subsequent chemical steps. In Step2 anilide amide hydrolysis under standard acidic conditions maybeconducted followed by methyl ester synthesis under standard conditionsfor example as shown with methyl iodide and base gives correspondingmethyl 2-chloro-3-substituted isonicotinates. In Step 4 an alkylaminogroup can be introduced by Buchwald coupling reaction of an R₇NH₂monoalkylamine with the methyl 2-chloro-3-substituted isonicotinates.This reaction is well precedented for diverse 2-chloropyridine systemsin the chemical literature. In an optional Step 5 for dialkylaminocompounds R₈ groups can be introduced by reductive amination withR₈-ketone or R₈-aldehyde in the presence of an appropriate reducingagent such as sodium cyanoborohydride and catalytic acid such as aceticacid in an appropriate solvent such as methanol. Alternatively, avariety of R₈ groups can be introduced by alkylation using R₈-LG, whereLG is a leaving group such as iodine, in the presence of a mild basesuch as cesium carbonate in an appropriate polar solvent such asacetonitrile at an appropriate temperature such as 80° C. In Step 6,oxidation to the N-oxide followed by chlorination with phosphorusoxychloride gives methyl 6-chloro-2-mono or dialkylamino-3-substitutedisonicotinates. In Step 7 the ester moiety can be hydrolyzed to thecorresponding acid using a suitable base such as sodium hydroxide in apolar solvent such as ethanol. In Step 8, the acid can be subjected to astandard amide coupling reaction whereupon the appropriate amine orsubstituted 3-(aminomethyl)-pyridin-2(1H)-one would be added along witha suitable amide coupling reagent such as PYBOP in a suitable solventsuch as DMSO to give the desired amide. In Step 9, aryl groupscorresponding to R₆ can be introduced by Suzuki reaction of theintermediate bromide with an appropriate aryl boronic acid or esterderivative, e.g., X—Ar—B(OH)₂, in the presence of a mild base and apalladium catalyst in a polar solvent such as dioxane/water, at elevatedtemperature. The X group in X—Ar—B(OH)₂ may be a fully elaboratedsubstituent on the aryl ring or may be a functional group that can beconverted into another group by functional group modification. Arepresentative sampling of such modifications could includehydrogenation, protecting group removal followed by additional amidecoupling reactions, palladium catalyzed coupling reactions, reductiveamination reactions or alkylation reactions. For example if the Suzukireaction is conducted with a boronic acid derivative bearing a formylgroup further modification by reductive amination reaction with primaryand secondary amines (e.g. morpholine, dimethylamine) can be conductedto introduce amine groups. Depending upon the nature of the R₇substituent, further chemical modification steps may be employed toconvert the R₇ substituent into an alternative R₇ substituent. Forexample a protected amino group contained within R₇ may be subjected todeprotection reaction (e.g. Boc group cleavage) to give free aminogroups. Such free amino groups may be subjected to reductive aminationreactions or alkylation reactions to give substituted amines.

Scheme 9 depicts a synthesis of modified aryl analogs following ageneral route that utilizes well-established chemistry. Starting with asubstituted benzoic acid such as 5-chloro-2-methylbenzoic acid,nitration using standard conditions such as treatment with conc. H₂SO₄and conc. HNO₃ can provide the nitro analog. Esterification of the acidcan be achieved using an alkylating agent such as methyl iodide in thepresence of a base such as sodium carbonate in a polar solvent such asDMF. The nitro group can be reduced using conditions such iron andammonium chloride in a protic solvent such as ethanol with heating to atemperature such as 80° C. The resulting aniline can be converted to abromide using a Sandmeyer reaction such treatment with CuBr₂ and t-butylnitrite in a solvent such as acetonitrile. A palladium catalyzedcoupling of a thiol with the bromide can be achieved using a palladiumsource such as Pd(OAc)₂ with a ligand such as Xanthphos in the presenceof a base such as N,N-diisopropyl ethylamine in a solvent such as1,4-dioxane optionally heating to a temperature such as 100° C. Theester can be hydrolyzed with an aqueous base such as NaOH in water. Theresulting acid can be coupled to the 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one using standard amino acid couplingconditions such as PyBOP in DMSO. The resulting thioether may beoxidized to the corresponding sulfoxide or sulfone by using theappropriate equivalents of an oxidant such as m-CPBA in a solvent suchas DCM. Aryl substituents can be incorporated by using palladiumcouplings such as a Suzuki reaction as described above.

Scheme 10 depicts a synthesis of modified aryl analogs following ageneral route that utilizes well-established chemistry. Starting with asubstituted aniline such as methyl 3-amino-5-chloro-2-methylbenzoate,the aniline can be converted to a phenol using a Sandmeyer reaction suchas treatment with aqueous NaNO₂ solution in a aqueous acid such as 50%H₂SO₄. The phenol can be alkylated using an alkylating agent such astetrahydro-2H-pyran-4-yl 4-methylbenzenesulfonate in the presence of anappropriate base such as cesium carbonate in as polar solvent such asDMF optionally heating to a temperature such as 80° C. The ester can behydrolyzed with an aqueous base such as NaOH in water. The resultingacid can be coupled to the 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one using standard amino acid couplingconditions such as PyBOP in DMSO. Aryl substituents can be incorporatedby using palladium couplings such as a Suzuki reaction as describedabove.

3. METHODS OF TREATMENT

Compounds of the present invention inhibit the histone methyltransferaseactivity of EZH2 or a mutant thereof and, accordingly, in one aspect ofthe invention, certain compounds disclosed herein are candidates fortreating, or preventing certain conditions and diseases. The presentinvention provides methods for treating conditions and diseases thecourse of which can be influenced by modulating the methylation statusof histones or other proteins, wherein said methylation status ismediated at least in part by the activity of EZH2. Modulation of themethylation status of histones can in turn influence the level ofexpression of target genes activated by methylation, and/or target genessuppressed by methylation. The method includes administering to asubject in need of such treatment, a therapeutically effective amount ofa compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph, solvate, or stereoisomerorthereof.

The disorder in which EZH2-mediated protein methylation plays a part canbe cancer or a precancerous condition. The present invention furtherprovides the use of a compound of the present invention, or apharmaceutically acceptable salt, ester, prodrug, metabolite, polymorphor solvate thereof in the treatment of cancer or precancer the course ofwhich can be influenced by modulating EZH2-mediated protein methylation,or, for the preparation of a medicament useful for the treatment of suchcancer or pre-cancer. Exemplary cancers that may be treated includelymphomas, including non-Hodgkin lymphoma, follicular lymphoma (FL) anddiffuse large B-cell lymphoma (DLBCL); melanoma; and leukemia, includingCML. Exemplary precancerous condition includes myelodysplastic syndrome(MDS; formerly known as preleukemia).

The present invention also provides methods of protecting against adisorder in which EZH2-mediated protein methylation plays a part in asubject in need thereof by administering a therapeutically effectiveamount of compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, polymorph or solvate thereof, to asubject in need of such treatment. The disorder can be cancer, e.g.,cancer in which EZH2-mediated protein methylation plays a role. Thepresent invention also provides the use of compound of the presentinvention, or a pharmaceutically acceptable salt, ester, prodrug,metabolite, polymorph, solvate, or stereoisomeror thereof, for thepreparation of a medicament useful for the prevention of a cellproliferative disorder associated, at least in part, with EZH2-mediatedprotein methylation.

The compounds of this invention can be used to modulate protein (e.g.,histone) methylation, e.g., to modulate histone methyltransferase orhistone demethylase enzyme activity. At least some of the compounds ofthe invention can be used in vivo or in vitro for modulating proteinmethylation. Histone methylation has been reported to be involved inaberrant expression of certain genes in cancers, and in silencing ofneuronal genes in non-neuronal cells. At least some compounds describedherein are suitable candidates for treating these diseases, i.e., todecrease methylation or restore methylation to roughly its level incounterpart normal cells.

Compounds that are methylation modulators may be used for modulatingcell proliferation. For example, in some cases excessive proliferationmay be reduced with agents that decrease methylation, whereasinsufficient proliferation may be stimulated with agents that increasemethylation. Accordingly, diseases that may be treated by the compoundsof the invention can include hyperproliferative diseases, such as benigncell growth and malignant cell growth.

As used herein, a “subject in need thereof” is a subject having adisorder in which EZH2-mediated protein methylation plays a part, or asubject having an increased risk of developing such disorder relative tothe population at large. A subject in need thereof can have aprecancerous condition. Preferably, a subject in need thereof hascancer. A “subject” includes a mammal. The mammal can be e.g., a humanor appropriate non-human mammal, such as primate, mouse, rat, dog, cat,cow, horse, goat, camel, sheep or a pig. The subject can also be a birdor fowl. In one embodiment, the mammal is a human.

As used herein, the term “cell proliferative disorder” refers toconditions in which unregulated or abnormal growth, or both, of cellscan lead to the development of an unwanted condition or disease, whichmay or may not be cancerous. Exemplary cell proliferative disorders thatmay be treated with the compounds of the invention encompass a varietyof conditions wherein cell division is deregulated. Exemplary cellproliferative disorder include, but are not limited to, neoplasms,benign tumors, malignant tumors, pre-cancerous conditions, in situtumors, encapsulated tumors, metastatic tumors, liquid tumors, solidtumors, immunological tumors, hematological tumors, cancers, carcinomas,leukemias, lymphomas, sarcomas, and rapidly dividing cells. The term“rapidly dividing cell” as used herein is defined as any cell thatdivides at a rate that exceeds or is greater than what is expected orobserved among neighboring or juxtaposed cells within the same tissue. Acell proliferative disorder includes a precancer or a precancerouscondition. A cell proliferative disorder includes cancer. In one aspect,the methods provided herein are used to treat or alleviate a symptom ofcancer or to identify suitable candidates for such purposes. The term“cancer” includes solid tumors, as well as, hematologic tumors and/ormalignancies. A “precancer cell” or “precancerous cell” is a cellmanifesting a cell proliferative disorder that is a precancer or aprecancerous condition. A “cancer cell” or “cancerous cell” is a cellmanifesting a cell proliferative disorder that is a cancer. Anyreproducible means of measurement may be used to identify cancer cellsor precancerous cells. Cancer cells or precancerous cells can beidentified by histological typing or grading of a tissue sample (e.g., abiopsy sample). Cancer cells or precancerous cells can be identifiedthrough the use of appropriate molecular markers.

Exemplary non-cancerous conditions or disorders that may be treatedusing one or more compounds of the present invention include, but arenot limited to, rheumatoid arthritis; inflammation; autoimmune disease;lymphoproliferative conditions; acromegaly; rheumatoid spondylitis;osteoarthritis; gout, other arthritic conditions; sepsis; septic shock;endotoxic shock; gram-negative sepsis; toxic shock syndrome; asthma;adult respiratory distress syndrome; chronic obstructive pulmonarydisease; chronic pulmonary inflammation; inflammatory bowel disease;Crohn's disease; psoriasis; eczema; ulcerative colitis; pancreaticfibrosis; hepatic fibrosis; acute and chronic renal disease; irritablebowel syndrome; pyresis; restenosis; cerebral malaria; stroke andischemic injury; neural trauma; Alzheimer's disease; Huntington'sdisease; Parkinson's disease; acute and chronic pain; allergic rhinitis;allergic conjunctivitis; chronic heart failure; acute coronary syndrome;cachexia; malaria; leprosy; leishmaniasis; Lyme disease; Reiter'ssyndrome; acute synovitis; muscle degeneration, bursitis; tendonitis;tenosynovitis; herniated, ruptures, or prolapsed intervertebral disksyndrome; osteopetrosis; thrombosis; restenosis; silicosis; pulmonarysarcosis; bone resorption diseases, such as osteoporosis;graft-versus-host reaction; Multiple Sclerosis; lupus; fibromyalgia;AIDS and other viral diseases such as Herpes Zoster, Herpes Simplex I orII, influenza virus and cytomegalovirus; and diabetes mellitus.

Exemplary cancers that may be treated using one or more compounds of thepresent invention include, but are not limited to, adrenocorticalcarcinoma, AIDS-related cancers, AIDS-related lymphoma, anal cancer,anorectal cancer, cancer of the anal canal, appendix cancer, childhoodcerebellar astrocytoma, childhood cerebral astrocytoma, basal cellcarcinoma, skin cancer (non-melanoma), biliary cancer, extrahepatic bileduct cancer, intrahepatic bile duct cancer, bladder cancer, urinarybladder cancer, bone and joint cancer, osteosarcoma and malignantfibrous histiocytoma, brain cancer, brain tumor, brain stem glioma,cerebellar astrocytoma, cerebral astrocytoma/malignant glioma,ependymoma, medulloblastoma, supratentorial primitive neuroectodeimaltumors, visual pathway and hypothalamic glioma, breast cancer, bronchialadenomas/carcinoids, carcinoid tumor, gastrointestinal, nervous systemcancer, nervous system lymphoma, central nervous system cancer, centralnervous system lymphoma, cervical cancer, childhood cancers, chroniclymphocytic leukemia, chronic myelogenous leukemia, chronicmyeloproliferative disorders, colon cancer, colorectal cancer, cutaneousT-cell lymphoma, lymphoid neoplasm, mycosis fungoides, Seziary Syndrome,endometrial cancer, esophageal cancer, extracranial germ cell tumor,extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer,intraocular melanoma, retinoblastoma, gallbladder cancer, gastric(stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinalstromal tumor (GIST), germ cell tumor, ovarian germ cell tumor,gestational trophoblastic tumor glioma, head and neck cancer,hepatocellular (liver) cancer, Hodgkin lymphoma, hypopharyngeal cancer,intraocular melanoma, ocular cancer, islet cell tumors (endocrinepancreas), Kaposi Sarcoma, kidney cancer, renal cancer, kidney cancer,laryngeal cancer, acute lymphoblastic leukemia, acute myeloid leukemia,chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cellleukemia, lip and oral cavity cancer, liver cancer, lung cancer,non-small cell lung cancer, small cell lung cancer, AIDS-relatedlymphoma, non-Hodgkin lymphoma, primary central nervous system lymphoma,Waldenstram macroglobulinemia, medulloblastoma, melanoma, intraocular(eye) melanoma, merkel cell carcinoma, mesothelioma malignant,mesothelioma, metastatic squamous neck cancer, mouth cancer, cancer ofthe tongue, multiple endocrine neoplasia syndrome, mycosis fungoides,myelodysplastic syndromes, myelodysplastic/myeloproliferative diseases,chronic myelogenous leukemia, acute myeloid leukemia, multiple myeloma,chronic myeloproliferative disorders, nasopharyngeal cancer,neuroblastoma, oral cancer, oral cavity cancer, oropharyngeal cancer,ovarian cancer, ovarian epithelial cancer, ovarian low malignantpotential tumor, pancreatic cancer, islet cell pancreatic cancer,paranasal sinus and nasal cavity cancer, parathyroid cancer, penilecancer, pharyngeal cancer, pheochromocytoma, pineoblastoma andsupratentorial primitive neuroectodermal tumors, pituitary tumor, plasmacell neoplasm/multiple myeloma, pleuropulmonary blastoma, prostatecancer, rectal cancer, renal pelvis and ureter, transitional cellcancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, ewingfamily of sarcoma tumors, Kaposi Sarcoma, soft tissue sarcoma, uterinecancer, uterine sarcoma, skin cancer (non-melanoma), skin cancer(melanoma), merkel cell skin carcinoma, small intestine cancer, softtissue sarcoma, squamous cell carcinoma, stomach (gastric) cancer,supratentorial primitive neuroectodermal tumors, testicular cancer,throat cancer, thymoma, thymoma and thymic carcinoma, thyroid cancer,transitional cell cancer of the renal pelvis and ureter and otherurinary organs, gestational trophoblastic tumor, urethral cancer,endometrial uterine cancer, uterine sarcoma, uterine corpus cancer,vaginal cancer, vulvar cancer, and Wilm's Tumor.

A “cell proliferative disorder of the hematologic system” is a cellproliferative disorder involving cells of the hematologic system. A cellproliferative disorder of the hematologic system can include lymphoma,leukemia, myeloid neoplasms, mast cell neoplasms, myelodysplasia, benignmonoclonal gammopathy, lymphomatoid granulomatosis, lymphomatoidpapulosis, polycythemia vera, chronic myelocytic leukemia, agnogenicmyeloid metaplasia, and essential thrombocythemia. A cell proliferativedisorder of the hematologic system can include hyperplasia, dysplasia,and metaplasia of cells of the hematologic system. In one aspect,compositions of the present invention may be used to treat a cancerselected from the group consisting of a hematologic cancer of thepresent invention or a hematologic cell proliferative disorder of thepresent invention, or used to identify suitable candidates for suchpurposes. A hematologic cancer of the present invention can includemultiple myeloma, lymphoma (including Hodgkin's lymphoma, non-Hodgkin'slymphoma, childhood lymphomas, and lymphomas of lymphocytic andcutaneous origin), leukemia (including childhood leukemia, hairy-cellleukemia, acute lymphocytic leukemia, acute myelocytic leukemia, chroniclymphocytic leukemia, chronic myelocytic leukemia, chronic myelogenousleukemia, and mast cell leukemia), myeloid neoplasms and mast cellneoplasms.

A “cell proliferative disorder of the lung” is a cell proliferativedisorder involving cells of the lung. Cell proliferative disorders ofthe lung can include all forms of cell proliferative disorders affectinglung cells. Cell proliferative disorders of the lung can include lungcancer, a precancer or precancerous condition of the lung, benigngrowths or lesions of the lung, and malignant growths or lesions of thelung, and metastatic lesions in tissue and organs in the body other thanthe lung. In one aspect, compositions of the present invention may beused to treat lung cancer or cell proliferative disorders of the lung,or used to identify suitable candidates for such purposes. Lung cancercan include all forms of cancer of the lung. Lung cancer can includemalignant lung neoplasms, carcinoma in situ, typical carcinoid tumors,and atypical carcinoid tumors. Lung cancer can include small cell lungcancer (“SCLC”), non-small cell lung cancer (“NSCLC”), squamous cellcarcinoma, adenocarcinoma, small cell carcinoma, large cell carcinoma,adenosquamous cell carcinoma, and mesothelioma. Lung cancer can include“scar carcinoma,” bronchioalveolar carcinoma, giant cell carcinoma,spindle cell carcinoma, and large cell neuroendocrine carcinoma. Lungcancer can include lung neoplasms having histologic and ultrastructualheterogeneity (e.g., mixed cell types).

Cell proliferative disorders of the lung can include all forms of cellproliferative disorders affecting lung cells. Cell proliferativedisorders of the lung can include lung cancer, precancerous conditionsof the lung. Cell proliferative disorders of the lung can includehyperplasia, metaplasia, and dysplasia of the lung. Cell proliferativedisorders of the lung can include asbestos-induced hyperplasia, squamousmetaplasia, and benign reactive mesothelial metaplasia. Cellproliferative disorders of the lung can include replacement of columnarepithelium with stratified squamous epithelium, and mucosal dysplasia.Individuals exposed to inhaled injurious environmental agents such ascigarette smoke and asbestos may be at increased risk for developingcell proliferative disorders of the lung. Prior lung diseases that maypredispose individuals to development of cell proliferative disorders ofthe lung can include chronic interstitial lung disease, necrotizingpulmonary disease, scleroderma, rheumatoid disease, sarcoidosis,interstitial pneumonitis, tuberculosis, repeated pneumonias, idiopathicpulmonary fibrosis, granulomata, asbestosis, fibrosing alveolitis, andHodgkin's disease.

A “cell proliferative disorder of the colon” is a cell proliferativedisorder involving cells of the colon. Preferably, the cellproliferative disorder of the colon is colon cancer. In one aspect,compositions of the present invention may be used to treat colon canceror cell proliferative disorders of the colon, or used to identifysuitable candidates for such purposes. Colon cancer can include allforms of cancer of the colon. Colon cancer can include sporadic andhereditary colon cancers. Colon cancer can include malignant colonneoplasms, carcinoma in situ, typical carcinoid tumors, and atypicalcarcinoid tumors. Colon cancer can include adenocarcinoma, squamous cellcarcinoma, and adenosquamous cell carcinoma. Colon cancer can beassociated with a hereditary syndrome selected from the group consistingof hereditary nonpolyposis colorectal cancer, familial adenomatouspolyposis, Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's syndromeand juvenile polyposis. Colon cancer can be caused by a hereditarysyndrome selected from the group consisting of hereditary nonpolyposiscolorectal cancer, familial adenomatous polyposis, Gardner's syndrome,Peutz-Jeghers syndrome, Turcot's syndrome and juvenile polyposis.

Cell proliferative disorders of the colon can include all forms of cellproliferative disorders affecting colon cells. Cell proliferativedisorders of the colon can include colon cancer, precancerous conditionsof the colon, adenomatous polyps of the colon and metachronous lesionsof the colon. A cell proliferative disorder of the colon can includeadenoma. Cell proliferative disorders of the colon can be characterizedby hyperplasia, metaplasia, and dysplasia of the colon. Prior colondiseases that may predispose individuals to development of cellproliferative disorders of the colon can include prior colon cancer.Current disease that may predispose individuals to development of cellproliferative disorders of the colon can include Crohn's disease andulcerative colitis. A cell proliferative disorder of the colon can beassociated with a mutation in a gene selected from the group consistingof p53, ras, FAP and DCC. An individual can have an elevated risk ofdeveloping a cell proliferative disorder of the colon due to thepresence of a mutation in a gene selected from the group consisting ofp53, ras, FAP and DCC.

A “cell proliferative disorder of the pancreas” is a cell proliferativedisorder involving cells of the pancreas. Cell proliferative disordersof the pancreas can include all forms of cell proliferative disordersaffecting pancreatic cells. Cell proliferative disorders of the pancreascan include pancreas cancer, a precancer or precancerous condition ofthe pancreas, hyperplasia of the pancreas, and dysaplasia of thepancreas, benign growths or lesions of the pancreas, and malignantgrowths or lesions of the pancreas, and metastatic lesions in tissue andorgans in the body other than the pancreas. Pancreatic cancer includesall forms of cancer of the pancreas. Pancreatic cancer can includeductal adenocarcinoma, adenosquamous carcinoma, pleomorphic giant cellcarcinoma, mucinous adenocarcinoma, osteoclast-like giant cellcarcinoma, mucinous cystadenocarcinoma, acinar carcinoma, unclassifiedlarge cell carcinoma, small cell carcinoma, pancreatoblastoma, papillaryneoplasm, mucinous cystadenoma, papillary cystic neoplasm, and serouscystadenoma. Pancreatic cancer can also include pancreatic neoplasmshaving histologic and ultrastructual heterogeneity (e.g., mixed celltypes).

A “cell proliferative disorder of the prostate” is a cell proliferativedisorder involving cells of the prostate. Cell proliferative disordersof the prostate can include all forms of cell proliferative disordersaffecting prostate cells. Cell proliferative disorders of the prostatecan include prostate cancer, a precancer or precancerous condition ofthe prostate, benign growths or lesions of the prostate, and malignantgrowths or lesions of the prostate, and metastatic lesions in tissue andorgans in the body other than the prostate. Cell proliferative disordersof the prostate can include hyperplasia, metaplasia, and dysplasia ofthe prostate.

A “cell proliferative disorder of the skin” is a cell proliferativedisorder involving cells of the skin. Cell proliferative disorders ofthe skin can include all forms of cell proliferative disorders affectingskin cells. Cell proliferative disorders of the skin can include aprecancer or precancerous condition of the skin, benign growths orlesions of the skin, melanoma, malignant melanoma and other malignantgrowths or lesions of the skin, and metastatic lesions in tissue andorgans in the body other than the skin. Cell proliferative disorders ofthe skin can include hyperplasia, metaplasia, and dysplasia of the skin.

A “cell proliferative disorder of the ovary” is a cell proliferativedisorder involving cells of the ovary. Cell proliferative disorders ofthe ovary can include all forms of cell proliferative disordersaffecting cells of the ovary. Cell proliferative disorders of the ovarycan include a precancer or precancerous condition of the ovary, benigngrowths or lesions of the ovary, ovarian cancer, malignant growths orlesions of the ovary, and metastatic lesions in tissue and organs in thebody other than the ovary. Cell proliferative disorders of the skin caninclude hyperplasia, metaplasia, and dysplasia of cells of the ovary.

A “cell proliferative disorder of the breast” is a cell proliferativedisorder involving cells of the breast. Cell proliferative disorders ofthe breast can include all forms of cell proliferative disordersaffecting breast cells. Cell proliferative disorders of the breast caninclude breast cancer, a precancer or precancerous condition of thebreast, benign growths or lesions of the breast, and malignant growthsor lesions of the breast, and metastatic lesions in tissue and organs inthe body other than the breast. Cell proliferative disorders of thebreast can include hyperplasia, metaplasia, and dysplasia of the breast.

A cell proliferative disorder of the breast can be a precancerouscondition of the breast. Compositions of the present invention may beused to treat a precancerous condition of the breast. A precancerouscondition of the breast can include atypical hyperplasia of the breast,ductal carcinoma in situ (DCIS), intraductal carcinoma, lobularcarcinoma in situ (LCIS), lobular neoplasia, and stage 0 or grade 0growth or lesion of the breast (e.g., stage 0 or grade 0 breast cancer,or carcinoma in situ). A precancerous condition of the breast can bestaged according to the TNM classification scheme as accepted by theAmerican Joint Committee on Cancer (AJCC), where the primary tumor (T)has been assigned a stage of T0 or Tis; and where the regional lymphnodes (N) have been assigned a stage of N0; and where distant metastasis(M) has been assigned a stage of M0.

The cell proliferative disorder of the breast can be breast cancer. Inone aspect, compositions of the present invention may be used to treatbreast cancer, or used to identify suitable candidates for suchpurposes. Breast cancer may include all forms of cancer of the breast.Breast cancer can include primary epithelial breast cancers. Breastcancer can include cancers in which the breast is involved by othertumors such as lymphoma, sarcoma or melanoma. Breast cancer can includecarcinoma of the breast, ductal carcinoma of the breast, lobularcarcinoma of the breast, undifferentiated carcinoma of the breast,cystosarcoma phyllodes of the breast, angiosarcoma of the breast, andprimary lymphoma of the breast. Breast cancer can include Stage I, II,IIIA, IIIB, IIIC and IV breast cancer. Ductal carcinoma of the breastcan include invasive carcinoma, invasive carcinoma in situ withpredominant intraductal component, inflammatory breast cancer, and aductal carcinoma of the breast with a histologic type selected from thegroup consisting of comedo, mucinous (colloid), medullary, medullarywith lymphcytic infiltrate, papillary, scirrhous, and tubular. Lobularcarcinoma of the breast can include invasive lobular carcinoma withpredominant in situ component, invasive lobular carcinoma, andinfiltrating lobular carcinoma. Breast cancer can include Paget'sdisease, Paget's disease with intraductal carcinoma, and Paget's diseasewith invasive ductal carcinoma. Breast cancer can include breastneoplasms having histologic and ultrastructual heterogeneity (e.g.,mixed cell types).

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph, or solvate thereof, may be used totreat breast cancer, or used to identify suitable candidates for suchpurposes. A breast cancer that is to be treated can include familialbreast cancer. A breast cancer that is to be treated can includesporadic breast cancer. A breast cancer that is to be treated can arisein a male subject. A breast cancer that is to be treated can arise in afemale subject. A breast cancer that is to be treated can arise in apremenopausal female subject or a postmenopausal female subject. Abreast cancer that is to be treated can arise in a subject equal to orolder than 30 years old, or a subject younger than 30 years old. Abreast cancer that is to be treated has arisen in a subject equal to orolder than 50 years old, or a subject younger than 50 years old. Abreast cancer that is to be treated can arise in a subject equal to orolder than 70 years old, or a subject younger than 70 years old.

A breast cancer that is to be treated can be typed to identify afamilial or spontaneous mutation in BRCA1, BRCA2, or p53. A breastcancer that is to be treated can be typed as having a HER2/neu geneamplification, as overexpressing HER2/neu, or as having a low,intermediate or high level of HER2/neu expression. A breast cancer thatis to be treated can be typed for a marker selected from the groupconsisting of estrogen receptor (ER), progesterone receptor (PR), humanepidermal growth factor receptor-2, Ki-67, CA15-3, CA 27-29, and c-Met.A breast cancer that is to be treated can be typed as ER-unknown,ER-rich or ER-poor. A breast cancer that is to be treated can be typedas ER-negative or ER-positive. ER-typing of a breast cancer may beperformed by any reproducible means. ER-typing of a breast cancer may beperformed as set forth in Onkologie 27: 175-179 (2004). A breast cancerthat is to be treated can be typed as PR-unknown, PR-rich, or PR-poor. Abreast cancer that is to be treated can be typed as PR-negative orPR-positive. A breast cancer that is to be treated can be typed asreceptor positive or receptor negative. A breast cancer that is to betreated can be typed as being associated with elevated blood levels ofCA 15-3, or CA 27-29, or both.

A breast cancer that is to be treated can include a localized tumor ofthe breast. A breast cancer that is to be treated can include a tumor ofthe breast that is associated with a negative sentinel lymph node (SLN)biopsy. A breast cancer that is to be treated can include a tumor of thebreast that is associated with a positive sentinel lymph node (SLN)biopsy. A breast cancer that is to be treated can include a tumor of thebreast that is associated with one or more positive axillary lymphnodes, where the axillary lymph nodes have been staged by any applicablemethod. A breast cancer that is to be treated can include a tumor of thebreast that has been typed as having nodal negative status (e.g.,node-negative) or nodal positive status (e.g., node-positive). A breastcancer that is to be treated can include a tumor of the breast that hasmetastasized to other locations in the body. A breast cancer that is tobe treated can be classified as having metastasized to a locationselected from the group consisting of bone, lung, liver, or brain. Abreast cancer that is to be treated can be classified according to acharacteristic selected from the group consisting of metastatic,localized, regional, local-regional, locally advanced, distant,multicentric, bilateral, ipsilateral, contralateral, newly diagnosed,recurrent, and inoperable.

A compound of the present invention, or a pharmaceutically acceptablesalt, ester, prodrug, metabolite, polymorph or solvate thereof, may beused to treat or prevent a cell proliferative disorder of the breast, orto treat or prevent breast cancer, in a subject having an increased riskof developing breast cancer relative to the population at large, or usedto identify suitable candidates for such purposes. A subject with anincreased risk of developing breast cancer relative to the population atlarge is a female subject with a family history or personal history ofbreast cancer. A subject with an increased risk of developing breastcancer relative to the population at large is a female subject having agerm-line or spontaneous mutation in BRCA1 or BRCA2, or both. A subjectwith an increased risk of developing breast cancer relative to thepopulation at large is a female subject with a family history of breastcancer and a germ-line or spontaneous mutation in BRCA1 or BRCA2, orboth. A subject with an increased risk of developing breast cancerrelative to the population at large is a female who is greater than 30years old, greater than 40 years old, greater than 50 years old, greaterthan 60 years old, greater than 70 years old, greater than 80 years old,or greater than 90 years old. A subject with an increased risk ofdeveloping breast cancer relative to the population at large is asubject with atypical hyperplasia of the breast, ductal carcinoma insitu (DCIS), intraductal carcinoma, lobular carcinoma in situ (LCIS),lobular neoplasia, or a stage 0 growth or lesion of the breast (e.g.,stage 0 or grade 0 breast cancer, or carcinoma in situ).

A breast cancer that is to be treated can histologically gradedaccording to the Scarff-Bloom-Richardson system, wherein a breast tumorhas been assigned a mitosis count score of 1, 2, or 3; a nuclearpleiomorphism score of 1, 2, or 3; a tubule formation score of 1, 2, or3; and a total Scarff-Bloom-Richardson score of between 3 and 9. Abreast cancer that is to be treated can be assigned a tumor gradeaccording to the International Consensus Panel on the Treatment ofBreast Cancer selected from the group consisting of grade 1, grade 1-2,grade 2, grade 2-3, or grade 3.

A cancer that is to be treated can be staged according to the AmericanJoint Committee on Cancer (AJCC) TNM classification system, where thetumor (T) has been assigned a stage of TX, T1, T1mic, T1a, T1b, T1c, T2,T3, T4, T4a, T4b, T4c, or T4d; and where the regional lymph nodes (N)have been assigned a stage of NX, N0, N1, N2, N2a, N2b, N3, N3a, N3b, orN3c; and where distant metastasis (M) can be assigned a stage of MX, M0,or M1. A cancer that is to be treated can be staged according to anAmerican Joint Committee on Cancer (AJCC) classification as Stage I,Stage IIA, Stage IIB, Stage IIIA, Stage IIIB, Stage IIIC, or Stage IV. Acancer that is to be treated can be assigned a grade according to anAJCC classification as Grade GX (e.g., grade cannot be assessed), Grade1, Grade 2, Grade 3 or Grade 4. A cancer that is to be treated can bestaged according to an AJCC pathologic classification (pN) of pNX, pN0,PN0 (I−), PN0 (I+), PN0 (mol−), PN0 (mol+), PN1, PN1(mi), PN1a, PN1b,PN1c, pN2, pN2a, pN2b, pN3, pN3a, pN3b, or pN3c.

A cancer that is to be treated can include a tumor that has beendetermined to be less than or equal to about 2 centimeters in diameter.A cancer that is to be treated can include a tumor that has beendetermined to be from about 2 to about 5 centimeters in diameter. Acancer that is to be treated can include a tumor that has beendetermined to be greater than or equal to about 3 centimeters indiameter. A cancer that is to be treated can include a tumor that hasbeen determined to be greater than 5 centimeters in diameter. A cancerthat is to be treated can be classified by microscopic appearance aswell differentiated, moderately differentiated, poorly differentiated,or undifferentiated. A cancer that is to be treated can be classified bymicroscopic appearance with respect to mitosis count (e.g., amount ofcell division) or nuclear pleiomorphism (e.g., change in cells). Acancer that is to be treated can be classified by microscopic appearanceas being associated with areas of necrosis (e.g., areas of dying ordegenerating cells). A cancer that is to be treated can be classified ashaving an abnormal karyotype, having an abnormal number of chromosomes,or having one or more chromosomes that are abnormal in appearance. Acancer that is to be treated can be classified as being aneuploid,triploid, tetraploid, or as having an altered ploidy. A cancer that isto be treated can be classified as having a chromosomal translocation,or a deletion or duplication of an entire chromosome, or a region ofdeletion, duplication or amplification of a portion of a chromosome.

A cancer that is to be treated can be evaluated by DNA cytometry, flowcytometry, or image cytometry. A cancer that is to be treated can betyped as having 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of cellsin the synthesis stage of cell division (e.g., in S phase of celldivision). A cancer that is to be treated can be typed as having a lowS-phase fraction or a high S-phase fraction.

As used herein, a “normal cell” is a cell that cannot be classified aspart of a “cell proliferative disorder”. A normal cell lacks unregulatedor abnormal growth, or both, that can lead to the development of anunwanted condition or disease. Preferably, a normal cell possessesnormally functioning cell cycle checkpoint control mechanisms.

As used herein, “contacting a cell” refers to a condition in which acompound or other composition of matter is in direct contact with acell, or is close enough to induce a desired biological effect in acell.

As used herein, “candidate compound” refers to a compound of the presentinvention, or a pharmaceutically acceptable salt, ester, prodrug,metabolite, polymorph or solvate thereof, that has been or will betested in one or more in vitro or in vivo biological assays, in order todetermine if that compound is likely to elicit a desired biological ormedical response in a cell, tissue, system, animal or human that isbeing sought by a researcher or clinician. A candidate compound is acompound of the present invention, or a pharmaceutically acceptablesalt, ester, prodrug, metabolite, polymorph or solvate thereof. Thebiological or medical response can be the treatment of cancer. Thebiological or medical response can be treatment or prevention of a cellproliferative disorder. The biological response or effect can alsoinclude a change in cell proliferation or growth that occurs in vitro orin an animal model, as well as other biological changes that areobservable in vitro. In vitro or in vivo biological assays can include,but are not limited to, enzymatic activity assays, electrophoreticmobility shift assays, reporter gene assays, in vitro cell viabilityassays, and the assays described herein.

As used herein, “monotherapy” refers to the administration of a singleactive or therapeutic compound to a subject in need thereof. Preferably,monotherapy will involve administration of a therapeutically effectiveamount of an active compound. For example, cancer monotherapy with oneof the compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, analog or derivative thereof, to asubject in need of treatment of cancer. Monotherapy may be contrastedwith combination therapy, in which a combination of multiple activecompounds is administered, preferably with each component of thecombination present in a therapeutically effective amount. In oneaspect, monotherapy with a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, is more effective than combination therapy in inducinga desired biological effect.

As used herein, “treating” or “treat” describes the management and careof a patient for the purpose of combating a disease, condition, ordisorder and includes the administration of a compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,polymorph or solvate thereof, to alleviate the symptoms or complicationsof a disease, condition or disorder, or to eliminate the disease,condition or disorder. The term “treat” can also include treatment of acell in vitro or an animal model.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, can also beused to prevent a disease, condition or disorder, or used to identifysuitable candidates for such purposes. As used herein, “preventing” or“prevent” describes reducing or eliminating the onset of the symptoms orcomplications of the disease, condition or disorder.

As used herein, the term “alleviate” is meant to describe a process bywhich the severity of a sign or symptom of a disorder is decreased.Importantly, a sign or symptom can be alleviated without beingeliminated. In a preferred embodiment, the administration ofpharmaceutical compositions of the invention leads to the elimination ofa sign or symptom, however, elimination is not required. Effectivedosages are expected to decrease the severity of a sign or symptom. Forinstance, a sign or symptom of a disorder such as cancer, which canoccur in multiple locations, is alleviated if the severity of the canceris decreased within at least one of multiple locations.

As used herein, the term “severity” is meant to describe the potentialof cancer to transform from a precancerous, or benign, state into amalignant state. Alternatively, or in addition, severity is meant todescribe a cancer stage, for example, according to the TNM system(accepted by the International Union Against Cancer (UICC) and theAmerican Joint Committee on Cancer (AJCC)) or by other art-recognizedmethods. Cancer stage refers to the extent or severity of the cancer,based on factors such as the location of the primary tumor, tumor size,number of tumors, and lymph node involvement (spread of cancer intolymph nodes). Alternatively, or in addition, severity is meant todescribe the tumor grade by art-recognized methods (see, National CancerInstitute, www.cancer.gov). Tumor grade is a system used to classifycancer cells in terms of how abnormal they look under a microscope andhow quickly the tumor is likely to grow and spread. Many factors areconsidered when determining tumor grade, including the structure andgrowth pattern of the cells. The specific factors used to determinetumor grade vary with each type of cancer. Severity also describes ahistologic grade, also called differentiation, which refers to how muchthe tumor cells resemble normal cells of the same tissue type (see,National Cancer Institute, www.cancer.gov). Furthermore, severitydescribes a nuclear grade, which refers to the size and shape of thenucleus in tumor cells and the percentage of tumor cells that aredividing (see, National Cancer Institute, www.cancer.gov).

In another aspect of the invention, severity describes the degree towhich a tumor has secreted growth factors, degraded the extracellularmatrix, become vascularized, lost adhesion to juxtaposed tissues, ormetastasized. Moreover, severity describes the number of locations towhich a primary tumor has metastasized. Finally, severity includes thedifficulty of treating tumors of varying types and locations. Forexample, inoperable tumors, those cancers which have greater access tomultiple body systems (hematological and immunological tumors), andthose which are the most resistant to traditional treatments areconsidered most severe. In these situations, prolonging the lifeexpectancy of the subject and/or reducing pain, decreasing theproportion of cancerous cells or restricting cells to one system, andimproving cancer stage/tumor grade/histological grade/nuclear grade areconsidered alleviating a sign or symptom of the cancer.

As used herein the term “symptom” is defined as an indication ofdisease, illness, injury, or that something is not right in the body.Symptoms are felt or noticed by the individual experiencing the symptom,but may not easily be noticed by others. Others are defined asnon-health-care professionals.

As used herein the term “sign” is also defined as an indication thatsomething is not right in the body. But signs are defined as things thatcan be seen by a doctor, nurse, or other health care professional.

Cancer is a group of diseases that may cause almost any sign or symptom.The signs and symptoms will depend on where the cancer is, the size ofthe cancer, and how much it affects the nearby organs or structures. Ifa cancer spreads (metastasizes), then symptoms may appear in differentparts of the body.

As a cancer grows, it begins to push on nearby organs, blood vessels,and nerves. This pressure creates some of the signs and symptoms ofcancer. If the cancer is in a critical area, such as certain parts ofthe brain, even the smallest tumor can cause early symptoms.

But sometimes cancers start in places where it does not cause anysymptoms until the cancer has grown quite large. Pancreas cancers, forexample, do not usually grow large enough to be felt from the outside ofthe body. Some pancreatic cancers do not cause symptoms until they beginto grow around nearby nerves (this causes a backache). Others growaround the bile duct, which blocks the flow of bile and leads to ayellowing of the skin known as jaundice. By the time a pancreatic cancercauses these signs or symptoms, it has usually reached an advancedstage.

A cancer may also cause symptoms such as fever, fatigue, or weight loss.This may be because cancer cells use up much of the body's energy supplyor release substances that change the body's metabolism. Or the cancermay cause the immune system to react in ways that produce thesesymptoms.

Sometimes, cancer cells release substances into the bloodstream thatcause symptoms not usually thought to result from cancers. For example,some cancers of the pancreas can release substances which cause bloodclots to develop in veins of the legs. Some lung cancers makehormone-like substances that affect blood calcium levels, affectingnerves and muscles and causing weakness and dizziness.

Cancer presents several general signs or symptoms that occur when avariety of subtypes of cancer cells are present. Most people with cancerwill lose weight at some time with their disease. An unexplained(unintentional) weight loss of 10 pounds or more may be the first signof cancer, particularly cancers of the pancreas, stomach, esophagus, orlung.

Fever is very common with cancer, but is more often seen in advanceddisease. Almost all patients with cancer will have fever at some time,especially if the cancer or its treatment affects the immune system andmakes it harder for the body to fight infection. Less often, fever maybe an early sign of cancer, such as with leukemia or lymphoma.

Fatigue may be an important symptom as cancer progresses. It may happenearly, though, in cancers such as with leukemia, or if the cancer iscausing an ongoing loss of blood, as in some colon or stomach cancers.

Pain may be an early symptom with some cancers such as bone cancers ortesticular cancer. But most often pain is a symptom of advanced disease.

Along with cancers of the skin (see next section), some internal cancerscan cause skin signs that can be seen. These changes include the skinlooking darker (hyperpigmentation), yellow (jaundice), or red(erythema); itching; or excessive hair growth.

Alternatively, or in addition, cancer subtypes present specific signs orsymptoms. Changes in bowel habits or bladder function could indicatecancer. Long-term constipation, diarrhea, or a change in the size of thestool may be a sign of colon cancer. Pain with urination, blood in theurine, or a change in bladder function (such as more frequent or lessfrequent urination) could be related to bladder or prostate cancer.

Changes in skin condition or appearance of a new skin condition couldindicate cancer. Skin cancers may bleed and look like sores that do notheal. A long-lasting sore in the mouth could be an oral cancer,especially in patients who smoke, chew tobacco, or frequently drinkalcohol. Sores on the penis or vagina may either be signs of infectionor an early cancer.

Unusual bleeding or discharge could indicate cancer. Unusual bleedingcan happen in either early or advanced cancer. Blood in the sputum(phlegm) may be a sign of lung cancer. Blood in the stool (or a dark orblack stool) could be a sign of colon or rectal cancer. Cancer of thecervix or the endometrium (lining of the uterus) can cause vaginalbleeding. Blood in the urine may be a sign of bladder or kidney cancer.A bloody discharge from the nipple may be a sign of breast cancer.

A thickening or lump in the breast or in other parts of the body couldindicate the presence of a cancer. Many cancers can be felt through theskin, mostly in the breast, testicle, lymph nodes (glands), and the softtissues of the body. A lump or thickening may be an early or late signof cancer. Any lump or thickening could be indicative of cancer,especially if the formation is new or has grown in size.

Indigestion or trouble swallowing could indicate cancer. While thesesymptoms commonly have other causes, indigestion or swallowing problemsmay be a sign of cancer of the esophagus, stomach, or pharynx (throat).

Recent changes in a wart or mole could be indicative of cancer. Anywart, mole, or freckle that changes in color, size, or shape, or losesits definite borders indicates the potential development of cancer. Forexample, the skin lesion may be a melanoma.

A persistent cough or hoarseness could be indicative of cancer. A coughthat does not go away may be a sign of lung cancer. Hoarseness can be asign of cancer of the larynx (voice box) or thyroid.

While the signs and symptoms listed above are the more common ones seenwith cancer, there are many others that are less common and are notlisted here. However, all art-recognized signs and symptoms of cancerare contemplated and encompassed by the instant invention.

Treating cancer can result in a reduction in size of a tumor. Areduction in size of a tumor may also be referred to as “tumorregression”. Preferably, after treatment, tumor size is reduced by 5% orgreater relative to its size prior to treatment; more preferably, tumorsize is reduced by 10% or greater; more preferably, reduced by 20% orgreater; more preferably, reduced by 30% or greater; more preferably,reduced by 40% or greater; even more preferably, reduced by 50% orgreater; and most preferably, reduced by greater than 75% or greater.Size of a tumor may be measured by any reproducible means ofmeasurement. The size of a tumor may be measured as a diameter of thetumor.

Treating cancer can result in a reduction in tumor volume. Preferably,after treatment, tumor volume is reduced by 5% or greater relative toits size prior to treatment; more preferably, tumor volume is reduced by10% or greater; more preferably, reduced by 20% or greater; morepreferably, reduced by 30% or greater; more preferably, reduced by 40%or greater; even more preferably, reduced by 50% or greater; and mostpreferably, reduced by greater than 75% or greater. Tumor volume may bemeasured by any reproducible means of measurement.

Treating cancer results in a decrease in number of tumors. Preferably,after treatment, tumor number is reduced by 5% or greater relative tonumber prior to treatment; more preferably, tumor number is reduced by10% or greater; more preferably, reduced by 20% or greater; morepreferably, reduced by 30% or greater; more preferably, reduced by 40%or greater; even more preferably, reduced by 50% or greater; and mostpreferably, reduced by greater than 75%. Number of tumors may bemeasured by any reproducible means of measurement. The number of tumorsmay be measured by counting tumors visible to the naked eye or at aspecified magnification. Preferably, the specified magnification is 2×,3×, 4×, 5×, 10×, or 50×.

Treating cancer can result in a decrease in number of metastatic lesionsin other tissues or organs distant from the primary tumor site.Preferably, after treatment, the number of metastatic lesions is reducedby 5% or greater relative to number prior to treatment; more preferably,the number of metastatic lesions is reduced by 10% or greater; morepreferably, reduced by 20% or greater; more preferably, reduced by 30%or greater; more preferably, reduced by 40% or greater; even morepreferably, reduced by 50% or greater; and most preferably, reduced bygreater than 75%. The number of metastatic lesions may be measured byany reproducible means of measurement. The number of metastatic lesionsmay be measured by counting metastatic lesions visible to the naked eyeor at a specified magnification. Preferably, the specified magnificationis 2×, 3×, 4×, 5×, 10×, or 50×.

Treating cancer can result in an increase in average survival time of apopulation of treated subjects in comparison to a population receivingcarrier alone. Preferably, the average survival time is increased bymore than 30 days; more preferably, by more than 60 days; morepreferably, by more than 90 days; and most preferably, by more than 120days. An increase in average survival time of a population may bemeasured by any reproducible means. An increase in average survival timeof a population may be measured, for example, by calculating for apopulation the average length of survival following initiation oftreatment with an active compound. An increase in average survival timeof a population may also be measured, for example, by calculating for apopulation the average length of survival following completion of afirst round of treatment with an active compound.

Treating cancer can result in an increase in average survival time of apopulation of treated subjects in comparison to a population ofuntreated subjects. Preferably, the average survival time is increasedby more than 30 days; more preferably, by more than 60 days; morepreferably, by more than 90 days; and most preferably, by more than 120days. An increase in average survival time of a population may bemeasured by any reproducible means. An increase in average survival timeof a population may be measured, for example, by calculating for apopulation the average length of survival following initiation oftreatment with an active compound. An increase in average survival timeof a population may also be measured, for example, by calculating for apopulation the average length of survival following completion of afirst round of treatment with an active compound.

Treating cancer can result in increase in average survival time of apopulation of treated subjects in comparison to a population receivingmonotherapy with a drug that is not a compound of the present invention,or a pharmaceutically acceptable salt, prodrug, metabolite, analog orderivative thereof. Preferably, the average survival time is increasedby more than 30 days; more preferably, by more than 60 days; morepreferably, by more than 90 days; and most preferably, by more than 120days. An increase in average survival time of a population may bemeasured by any reproducible means. An increase in average survival timeof a population may be measured, for example, by calculating for apopulation the average length of survival following initiation oftreatment with an active compound. An increase in average survival timeof a population may also be measured, for example, by calculating for apopulation the average length of survival following completion of afirst round of treatment with an active compound.

Treating cancer can result in a decrease in the mortality rate of apopulation of treated subjects in comparison to a population receivingcarrier alone. Treating cancer can result in a decrease in the mortalityrate of a population of treated subjects in comparison to an untreatedpopulation. Treating cancer can result in a decrease in the mortalityrate of a population of treated subjects in comparison to a populationreceiving monotherapy with a drug that is not a compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,analog or derivative thereof. Preferably, the mortality rate isdecreased by more than 2%; more preferably, by more than 5%; morepreferably, by more than 10%; and most preferably, by more than 25%. Adecrease in the mortality rate of a population of treated subjects maybe measured by any reproducible means. A decrease in the mortality rateof a population may be measured, for example, by calculating for apopulation the average number of disease-related deaths per unit timefollowing initiation of treatment with an active compound. A decrease inthe mortality rate of a population may also be measured, for example, bycalculating for a population the average number of disease-relateddeaths per unit time following completion of a first round of treatmentwith an active compound.

Treating cancer can result in a decrease in tumor growth rate.Preferably, after treatment, tumor growth rate is reduced by at least 5%relative to number prior to treatment; more preferably, tumor growthrate is reduced by at least 10%; more preferably, reduced by at least20%; more preferably, reduced by at least 30%; more preferably, reducedby at least 40%; more preferably, reduced by at least 50%; even morepreferably, reduced by at least 50%; and most preferably, reduced by atleast 75%. Tumor growth rate may be measured by any reproducible meansof measurement. Tumor growth rate can be measured according to a changein tumor diameter per unit time.

Treating cancer can result in a decrease in tumor regrowth. Preferably,after treatment, tumor regrowth is less than 5%; more preferably, tumorregrowth is less than 10%; more preferably, less than 20%; morepreferably, less than 30%; more preferably, less than 40%; morepreferably, less than 50%; even more preferably, less than 50%; and mostpreferably, less than 75%. Tumor regrowth may be measured by anyreproducible means of measurement. Tumor regrowth is measured, forexample, by measuring an increase in the diameter of a tumor after aprior tumor shrinkage that followed treatment. A decrease in tumorregrowth is indicated by failure of tumors to reoccur after treatmenthas stopped.

Treating or preventing a cell proliferative disorder can result in areduction in the rate of cellular proliferation. Preferably, aftertreatment, the rate of cellular proliferation is reduced by at least 5%;more preferably, by at least 10%; more preferably, by at least 20%; morepreferably, by at least 30%; more preferably, by at least 40%; morepreferably, by at least 50%; even more preferably, by at least 50%; andmost preferably, by at least 75%. The rate of cellular proliferation maybe measured by any reproducible means of measurement. The rate ofcellular proliferation is measured, for example, by measuring the numberof dividing cells in a tissue sample per unit time.

Treating or preventing a cell proliferative disorder can result in areduction in the proportion of proliferating cells. Preferably, aftertreatment, the proportion of proliferating cells is reduced by at least5%; more preferably, by at least 10%; more preferably, by at least 20%;more preferably, by at least 30%; more preferably, by at least 40%; morepreferably, by at least 50%; even more preferably, by at least 50%; andmost preferably, by at least 75%. The proportion of proliferating cellsmay be measured by any reproducible means of measurement. Preferably,the proportion of proliferating cells is measured, for example, byquantifying the number of dividing cells relative to the number ofnondividing cells in a tissue sample. The proportion of proliferatingcells can be equivalent to the mitotic index.

Treating or preventing a cell proliferative disorder can result in adecrease in size of an area or zone of cellular proliferation.Preferably, after treatment, size of an area or zone of cellularproliferation is reduced by at least 5% relative to its size prior totreatment; more preferably, reduced by at least 10%; more preferably,reduced by at least 20%; more preferably, reduced by at least 30%; morepreferably, reduced by at least 40%; more preferably, reduced by atleast 50%; even more preferably, reduced by at least 50%; and mostpreferably, reduced by at least 75%. Size of an area or zone of cellularproliferation may be measured by any reproducible means of measurement.The size of an area or zone of cellular proliferation may be measured asa diameter or width of an area or zone of cellular proliferation.

Treating or preventing a cell proliferative disorder can result in adecrease in the number or proportion of cells having an abnormalappearance or morphology. Preferably, after treatment, the number ofcells having an abnormal morphology is reduced by at least 5% relativeto its size prior to treatment; more preferably, reduced by at least10%; more preferably, reduced by at least 20%; more preferably, reducedby at least 30%; more preferably, reduced by at least 40%; morepreferably, reduced by at least 50%; even more preferably, reduced by atleast 50%; and most preferably, reduced by at least 75%. An abnormalcellular appearance or morphology may be measured by any reproduciblemeans of measurement. An abnormal cellular morphology can be measured bymicroscopy, e.g., using an inverted tissue culture microscope. Anabnormal cellular morphology can take the form of nuclear pleiomorphism.

As used herein, the term “selectively” means tending to occur at ahigher frequency in one population than in another population. Thecompared populations can be cell populations. Preferably, a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, acts selectively on a canceror precancerous cell but not on a normal cell. Preferably, a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, acts selectively to modulateone molecular target (e.g., a target protein methyltransferase) but doesnot significantly modulate another molecular target (e.g., a non-targetprotein methyltransferase). The invention also provides a method forselectively inhibiting the activity of an enzyme, such as a proteinmethyltransferase. Preferably, an event occurs selectively in populationA relative to population B if it occurs greater than two times morefrequently in population A as compared to population B. An event occursselectively if it occurs greater than five times more frequently inpopulation A. An event occurs selectively if it occurs greater than tentimes more frequently in population A; more preferably, greater thanfifty times; even more preferably, greater than 100 times; and mostpreferably, greater than 1000 times more frequently in population A ascompared to population B. For example, cell death would be said to occurselectively in cancer cells if it occurred greater than twice asfrequently in cancer cells as compared to normal cells.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, can modulatethe activity of a molecular target (e.g., a target proteinmethyltransferase). Modulating refers to stimulating or inhibiting anactivity of a molecular target. Preferably, a compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,polymorph or solvate thereof, modulates the activity of a moleculartarget if it stimulates or inhibits the activity of the molecular targetby at least 2-fold relative to the activity of the molecular targetunder the same conditions but lacking only the presence of saidcompound. More preferably, a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, modulates the activity of a molecular target if itstimulates or inhibits the activity of the molecular target by at least5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least100-fold relative to the activity of the molecular target under the sameconditions but lacking only the presence of said compound. The activityof a molecular target may be measured by any reproducible means. Theactivity of a molecular target may be measured in vitro or in vivo. Forexample, the activity of a molecular target may be measured in vitro byan enzymatic activity assay or a DNA binding assay, or the activity of amolecular target may be measured in vivo by assaying for expression of areporter gene.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, does notsignificantly modulate the activity of a molecular target if theaddition of the compound does not stimulate or inhibit the activity ofthe molecular target by greater than 10% relative to the activity of themolecular target under the same conditions but lacking only the presenceof said compound.

As used herein, the term “isozyme selective” means preferentialinhibition or stimulation of a first isoform of an enzyme in comparisonto a second isoform of an enzyme (e.g., preferential inhibition orstimulation of a protein methyltransferase isozyme alpha in comparisonto a protein methyltransferase isozyme beta). Preferably, a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, demonstrates a minimum of afourfold differential, preferably a tenfold differential, morepreferably a fifty fold differential, in the dosage required to achievea biological effect. Preferably, a compound of the present invention, ora pharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, demonstrates this differential across the range ofinhibition, and the differential is exemplified at the IC₅₀, i.e., a 50%inhibition, for a molecular target of interest.

Administering a compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, polymorph or solvate thereof, to acell or a subject in need thereof can result in modulation (i.e.,stimulation or inhibition) of an activity of a protein methyltransferaseof interest.

The present invention provides methods to assess biological activity ofa compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof or methods ofidentifying a test compound as an inhibitor of a Y641 mutant of EZH2. Inone embodiment the method includes combining an isolated Y641 mutant ofEZH2 with a histone substrate, a methyl group donor (such asS-adenosylmethionine (SAM)), and a test compound, wherein the histonesubstrate comprises a form of H3-K27 selected from the group consistingof unmethylated H3-K27, monomethylated H3-K27, dimethylated H3-K27, andany combination thereof, and performing an assay to detect methylationof H3-K27 in the histone substrate, thereby identifying the testcompound as an inhibitor of the Y641 mutant of EZH2 when methylation ofH3-K27 in the presence of the test compound is less than methylation ofH3-K27 in the absence of the test compound. The assay to detectmethylation of H3-K27 can be selected to measure the rate ofmethylation, the extent of methylation, or both the rate and extent ofmethylation.

The Y641 mutant of EZH2 is isolated as a PRC2 complex or functionalequivalent thereof. As used herein, the term “isolated” meanssubstantially separated from other components with which the complex maybe found as it occurs in nature. A compound can be isolated withoutnecessarily being purified. In one embodiment the mutant of EZH2 isisolated as a complex of a Y641 mutant of EZH2 together with EED andSUZ12. In another embodiment the mutant of EZH2 is isolated as a complexof a Y641 mutant of EZH2 together with EED, SUZ12, and RbAp48. Underappropriate conditions, a PRC2 complex or functional equivalent thereofexhibits histone methyltransferase activity for H3-K27. In oneembodiment the complex is composed of recombinantly expressed componentpolypeptides, e.g., EZH2, EED, SUZ12, with or without RbAp48.

The isolated Y641 mutant of EZH2 is combined with a histone substrate. Ahistone substrate includes any suitable source of histone polypeptidesor fragments thereof that can serve as substrate for EZH2. In oneembodiment the histone substrate includes histones isolated from asubject. The histones can be isolated from cells of a subject using anysuitable method; such methods are well known to persons skilled in theart and need not be further specified here. See, for example, Fang etal. (2004) Methods Enzymol 377:213-26. In accordance with the Examplesbelow, in one embodiment the histone substrate is provided asnucleosomes. In accordance with the Examples below, in one embodimentthe histone substrate is provided as avian (chicken) erythrocytenucleosomes.

Histone substrate so provided may include an admixture of states ofhistone modification, including various states of H3-K27 methylation asjudged by Western blotting with H3-K27 methylation state-specificantibodies. In one embodiment the histone substrate may be provided aspurified full-length histone H3. Such purified full-length histone H3may be provided as a homogeneous preparation in respect of states ofH3-K27 methylation, or as an admixture of various states of H3-K27methylation. Homogeneous preparations of isolated histone H3 in respectof states of H3-K27 methylation may be prepared in part by passage overan immunoaffinity column loaded with suitable H3-K27 methylationstate-specific antibodies or by immunoprecipitation using magnetic beadscoated with suitable H3-K27 methylation state-specific antibodies.Alternatively or in addition, the methylation state of H3-K27 can becharacterized as part of performing the assay. For example, the startingmaterial histone substrate might be characterized as containing 50percent unmethylated H3-K27, 40 percent monomethylated H3-K27, 10percent dimethylated H3-K27, and 0 percent trimethylated H3-K27.

In one embodiment the histone substrate includes a peptide library or asuitable peptide comprising one or more amino acid sequences related tohistone H3, including, in particular, a sequence that encompassesH3-K27. For example, in one embodiment, the histone substrate is apeptide fragment that corresponds to amino acid residues 21-44 ofhistone H3. The peptide library or peptide can be prepared by peptidesynthesis according to techniques well known in the art and optionallymodified so as to incorporate any desired degree of methylation oflysine corresponding to H3-K27. As described in the Examples below, suchpeptides can also be modified to incorporate a label, such as biotin,useful in performing downstream assays. In one embodiment the label isappended to the amino (N)-terminus of the peptide(s). In one embodimentthe label is appended to the carboxy (C)-terminus of the peptide(s).

Detection of methylation of H3-K27 can be accomplished using anysuitable method. In one embodiment, the source of donor methyl groupsincludes methyl groups that are labeled with a detectable label. Thedetectable label in one embodiment is an isotopic label, e.g., tritium.Other types of labels may include, for example, fluorescent labels.

Detection of formation of trimethylated H3-K27 can be accomplished usingany suitable method. For example, detection of formation oftrimethylated H3-K27 can be accomplished using an assay to detectincorporation of labeled methyl groups, such as described above,optionally combined with a chromatographic or other method to separatelabeled products by size, e.g., polyacrylamide gel electrophoresis(PAGE), capillary electrophoresis (CE), or high pressure liquidchromatography (HPLC). Alternatively or in addition, detection offormation of trimethylated H3-K27 can be accomplished using antibodiesthat are specific for trimethylated H3-K27.

Detection of conversion of monomethylated H3-K27 to dimethylated H3-K27can be accomplished using any suitable method. In one embodiment theconversion is measured using antibodies specific for monomethylatedH3-K27 and dimethylated H3-K27. For example, starting amounts orconcentrations of monomethylated H3-K27 and dimethylated H3-K27 may bedetermined using appropriate antibodies specific for monomethylatedH3-K27 and dimethylated H3-K27. Following the combination of enzyme,substrate, methyl group donor, and test compound, resulting amounts orconcentrations of monomethylated H3-K27 and dimethylated H3-K27 may thenbe determined using appropriate antibodies specific for monomethylatedH3-K27 and dimethylated H3-K27. The beginning and resulting amounts orconcentrations of monomethylated H3-K27 and dimethylated H3-K27 can thenbe compared. Alternatively or in addition, beginning and resultingamounts or concentrations of monomethylated H3-K27 and dimethylatedH3-K27 can then be compared to corresponding amounts of concentrationsfrom a negative control. A negative control reaction, in which no testagent is included in the assay, can be run in parallel or as ahistorical control. Results of such control reaction can optionally besubtracted from corresponding results of the experimental reaction priorto or in conjunction with making the comparison mentioned above.

Because the dimethylated form of H3-K27 may be further methylated in thesame assay, a reduction in the amount or concentration of monomethylatedH3-K27 may not appear to correspond directly to an increase indimethylated H3-K27. In this instance, it may be presumed, however, thata reduction in the amount or concentration of monomethylated H3-K27 is,by itself, reflective of conversion of monomethylated H3-K27 todimethylated H3-K27.

Detection of conversion of dimethylated H3-K27 to trimethylated H3-K27can be accomplished using any suitable method. In one embodiment theconversion is measured using antibodies specific for dimethylated H3-K27and trimethylated H3-K27. For example, starting amounts orconcentrations of dimethylated H3-K27 and trimethylated H3-K27 may bedetermined using appropriate antibodies specific for dimethylated H3-K27and trimethylated H3-K27. Following the combination of enzyme,substrate, and test compound, resulting amounts or concentrations ofdimethylated H3-K27 and trimethylated H3-K27 may then be determinedusing appropriate antibodies specific for dimethylated H3-K27 andtrimethylated H3-K27. The beginning and resulting amounts orconcentrations of dimethylated H3-K27 and trimethylated H3-K27 can thenbe compared. Alternatively or in addition, beginning and resultingamounts or concentrations of dimethylated H3-K27 and trimethylatedH3-K27 can then be compared to corresponding amounts of concentrationsfrom a negative control. A negative control reaction, in which no testagent is included in the assay, can be run in parallel or as ahistorical control. Results of such control reaction can optionally besubtracted from corresponding results of the experimental reaction priorto or in conjunction with making the comparison mentioned above.

A test agent is identified as an inhibitor of the Y641 mutant of EZH2when methylation of H3-K27 with the test compound is less thanmethylation of H3-K27 without the test compound. In one embodiment, atest agent is identified as an inhibitor of the Y641 mutant of EZH2 whenformation of trimethylated H3-K27 in the presence of the test compoundis less than formation of trimethylated H3-K27 in the absence of thetest compound.

The present invention also provides a method for identifying a selectiveinhibitor of a Y641 mutant of EZH2. In one embodiment the methodincludes combining an isolated Y641 mutant of EZH2 with a histonesubstrate, a methyl group donor (e.g., SAM), and a test compound,wherein the histone substrate comprises a form of H3-K27 selected fromthe group consisting of monomethylated H3-K27, dimethylated H3-K27, anda combination of monomethylated H3-K27 and dimethylated H3-K27, therebyforming a test mixture; combining an isolated wild-type EZH2 with ahistone substrate, a methyl group donor (e.g., SAM), and a testcompound, wherein the histone substrate comprises a form of H3-K27selected from the group consisting of monomethylated H3-K27,dimethylated H3-K27, and a combination of monomethylated H3-K27 anddimethylated H3-K27, thereby forming a control mixture; performing anassay to detect trimethylation of the histone substrate in each of thetest mixture and the control mixture; calculating the ratio of (a)trimethylation with the Y641 mutant of EZH2 and the test compound (M+)to (b) trimethylation with the Y641 mutant of EZH2 without the testcompound (M−); calculating the ratio of (c) trimethylation withwild-type EZH2 and the test compound (WT+) to (d) trimethylation withwild-type EZH2 without the test compound (WT−); comparing the ratio(a)/(b) with the ratio (c)/(d); and identifying the test compound as aselective inhibitor of the Y641 mutant of EZH2 when the ratio (a)/(b) isless than the ratio (c)/(d). In one embodiment the method furtherincludes taking into account a negative control without test compoundfor either or both of the test mixture and the control mixture.

In some assays, immunological reagents, e.g., antibodies and antigens,are employed. Fluorescence can be utilized in the measurement ofenzymatic activity in some assays. As used herein, “fluorescence” refersto a process through which a molecule emits a photon as a result ofabsorbing an incoming photon of higher energy by the same molecule.Specific methods for assessing the biological activity of the disclosedcompounds are described in the examples.

Administering a compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, polymorph or solvate thereof, to acell or a subject in need thereof results in modulation (i.e.,stimulation or inhibition) of an activity of an intracellular target(e.g., substrate). Several intracellular targets can be modulated withthe compounds of the present invention, including, but not limited to,protein methyltrasferase.

Activating refers to placing a composition of matter (e.g., protein ornucleic acid) in a state suitable for carrying out a desired biologicalfunction. A composition of matter capable of being activated also has anunactivated state. An activated composition of matter may have aninhibitory or stimulatory biological function, or both.

Elevation refers to an increase in a desired biological activity of acomposition of matter (e.g., a protein or a nucleic acid). Elevation mayoccur through an increase in concentration of a composition of matter.

As used herein, “a cell cycle checkpoint pathway” refers to abiochemical pathway that is involved in modulation of a cell cyclecheckpoint. A cell cycle checkpoint pathway may have stimulatory orinhibitory effects, or both, on one or more functions comprising a cellcycle checkpoint. A cell cycle checkpoint pathway is comprised of atleast two compositions of matter, preferably proteins, both of whichcontribute to modulation of a cell cycle checkpoint. A cell cyclecheckpoint pathway may be activated through an activation of one or moremembers of the cell cycle checkpoint pathway. Preferably, a cell cyclecheckpoint pathway is a biochemical signaling pathway.

As used herein, “cell cycle checkpoint regulator” refers to acomposition of matter that can function, at least in part, in modulationof a cell cycle checkpoint. A cell cycle checkpoint regulator may havestimulatory or inhibitory effects, or both, on one or more functionscomprising a cell cycle checkpoint. A cell cycle checkpoint regulatorcan be a protein or not a protein.

Treating cancer or a cell proliferative disorder can result in celldeath, and preferably, cell death results in a decrease of at least 10%in number of cells in a population. More preferably, cell death means adecrease of at least 20%; more preferably, a decrease of at least 30%;more preferably, a decrease of at least 40%; more preferably, a decreaseof at least 50%; most preferably, a decrease of at least 75%. Number ofcells in a population may be measured by any reproducible means. Anumber of cells in a population can be measured by fluorescenceactivated cell sorting (FACS), immunofluorescence microscopy and lightmicroscopy. Methods of measuring cell death are as shown in Li et al.,Proc Natl Acad Sci USA. 100(5): 2674-8, 2003. In an aspect, cell deathoccurs by apoptosis.

Preferably, an effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, is not significantly cytotoxic to normal cells. Atherapeutically effective amount of a compound is not significantlycytotoxic to normal cells if administration of the compound in atherapeutically effective amount does not induce cell death in greaterthan 10% of normal cells. A therapeutically effective amount of acompound does not significantly affect the viability of normal cells ifadministration of the compound in a therapeutically effective amountdoes not induce cell death in greater than 10% of normal cells. In anaspect, cell death occurs by apoptosis.

Contacting a cell with a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, can induce or activate cell death selectively in cancercells. Administering to a subject in need thereof a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, can induce or activate celldeath selectively in cancer cells. Contacting a cell with a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, can induce cell deathselectively in one or more cells affected by a cell proliferativedisorder. Preferably, administering to a subject in need thereof acompound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, induces celldeath selectively in one or more cells affected by a cell proliferativedisorder.

One aspect of the present invention relates to a method of treating orpreventing cancer (e.g., the course of which can be influenced bymodulating EZH2-mediated protein methylation) by administering acompound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, to a subject inneed thereof, where administration of the compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,polymorph or solvate thereof, results in one or more of the following:prevention of cancer cell proliferation by accumulation of cells in oneor more phases of the cell cycle (e.g. G1, G1/S, G2/M), or induction ofcell senescence, or promotion of tumor cell differentiation; promotionof cell death in cancer cells via cytotoxicity, necrosis or apoptosis,without a significant amount of cell death in normal cells, antitumoractivity in animals with a therapeutic index of at least 2. As usedherein, “therapeutic index” is the maximum tolerated dose divided by theefficacious dose. The present invention also relates to a method used toidentify suitable candidates for treating or preventing cancer.

One skilled in the art may refer to general reference texts for detaileddescriptions of known techniques discussed herein or equivalenttechniques. These texts include Ausubel et al., Current Protocols inMolecular Biology, John Wiley and Sons, Inc. (2005); Sambrook et al.,Molecular Cloning, A Laboratory Manual (3^(rd) edition), Cold SpringHarbor Press, Cold Spring Harbor, N.Y. (2000); Coligan et al., CurrentProtocols in Immunology, John Wiley & Sons, N.Y.; Enna et al., CurrentProtocols in Pharmacology, John Wiley & Sons, N.Y.; Fingl et al., ThePharmacological Basis of Therapeutics (1975), Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa., 18^(th) edition (1990).These texts can, of course, also be referred to in making or using anaspect of the invention.

As used herein, “combination therapy” or “co-therapy” includes theadministration of a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, and at least a second agent as part of a specifictreatment regimen intended to provide the beneficial effect from theco-action of these therapeutic agents. The beneficial effect of thecombination includes, but is not limited to, pharmacokinetic orpharmacodynamic co-action resulting from the combination of therapeuticagents. Administration of these therapeutic agents in combinationtypically is carried out over a defined time period (usually minutes,hours, days or weeks depending upon the combination selected).“Combination therapy” may be, but generally is not, intended toencompass the administration of two or more of these therapeutic agentsas part of separate monotherapy regimens that incidentally andarbitrarily result in the combinations of the present invention.

“Combination therapy” is intended to embrace administration of thesetherapeutic agents in a sequential manner, wherein each therapeuticagent is administered at a different time, as well as administration ofthese therapeutic agents, or at least two of the therapeutic agents, ina substantially simultaneous manner. Substantially simultaneousadministration can be accomplished, for example, by administering to thesubject a single capsule having a fixed ratio of each therapeutic agentor in multiple, single capsules for each of the therapeutic agents.Sequential or substantially simultaneous administration of eachtherapeutic agent can be effected by any appropriate route including,but not limited to, oral routes, intravenous routes, intramuscularroutes, and direct absorption through mucous membrane tissues. Thetherapeutic agents can be administered by the same route or by differentroutes. For example, a first therapeutic agent of the combinationselected may be administered by intravenous injection while the othertherapeutic agents of the combination may be administered orally.Alternatively, for example, all therapeutic agents may be administeredorally or all therapeutic agents may be administered by intravenousinjection. The sequence in which the therapeutic agents are administeredis not narrowly critical.

“Combination therapy” also embraces the administration of thetherapeutic agents as described above in further combination with otherbiologically active ingredients and non-drug therapies (e.g., surgery orradiation treatment). Where the combination therapy further comprises anon-drug treatment, the non-drug treatment may be conducted at anysuitable time so long as a beneficial effect from the co-action of thecombination of the therapeutic agents and non-drug treatment isachieved. For example, in appropriate cases, the beneficial effect isstill achieved when the non-drug treatment is temporally removed fromthe administration of the therapeutic agents, perhaps by days or evenweeks.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, analog or derivative thereof, may beadministered in combination with a second chemotherapeutic agent. Thesecond chemotherapeutic agent (also referred to as an anti-neoplasticagent or anti-proliferative agent) can be an alkylating agent; anantibiotic; an anti-metabolite; a detoxifying agent; an interferon; apolyclonal or monoclonal antibody; an EGFR inhibitor; a HER2 inhibitor;a histone deacetylase inhibitor; a hormone; a mitotic inhibitor; an MTORinhibitor; a multi-kinase inhibitor; a serine/threonine kinaseinhibitor; a tyrosine kinase inhibitors; a VEGF/VEGFR inhibitor; ataxane or taxane derivative, an aromatase inhibitor, an anthracycline, amicrotubule targeting drug, a topoisomerase poison drug, an inhibitor ofa molecular target or enzyme (e.g., a kinase or a proteinmethyltransferase), a cytidine analogue drug or any chemotherapeutic,anti-neoplastic or anti-proliferative agent listed inwww.cancer.org/docroot/cdg/cdg_0.asp.

Exemplary alkylating agents include, but are not limited to,cyclophosphamide (Cytoxan; Neosar); chlorambucil (Leukeran); melphalan(Alkeran); carmustine (BiCNU); busulfan (Busulfex); lomustine (CeeNU);dacarbazine (DTIC-Dome); oxaliplatin (Eloxatin); carmustine (Gliadel);ifosfamide (Ifex); mechlorethamine (Mustargen); busulfan (Myleran);carboplatin (Paraplatin); cisplatin (CDDP; Platinol); temozolomide(Temodar); thiotepa (Thioplex); bendamustine (Treanda); or streptozocin(Zanosar).

Exemplary antibiotics include, but are not limited to, doxorubicin(Adriamycin); doxorubicin liposomal (Doxil); mitoxantrone (Novantrone);bleomycin (Blenoxane); daunorubicin (Cerubidine); daunorubicin liposomal(DaunoXome); dactinomycin (Cosmegen); epirubicin (Ellence); idarubicin(Idamycin); plicamycin (Mithracin); mitomycin (Mutamycin); pentostatin(Nipent); or valrubicin (Valstar).

Exemplary anti-metabolites include, but are not limited to, fluorouracil(Adrucil); capecitabine (Xeloda); hydroxyurea (Hydrea); mercaptopurine(Purinethol); pemetrexed (Alimta); fludarabine (Fludara); nelarabine(Arranon); cladribine (Cladribine Novaplus); clofarabine (Clolar);cytarabine (Cytosar-U); decitabine (Dacogen); cytarabine liposomal(DepoCyt); hydroxyurea (Droxia); pralatrexate (Folotyn); floxuridine(FUDR); gemcitabine (Gemzar); cladribine (Leustatin); fludarabine(Oforta); methotrexate (MTX; Rheumatrex); methotrexate (Trexall);thioguanine (Tabloid); TS-1 or cytarabine (Tarabine PFS).

Exemplary detoxifying agents include, but are not limited to, amifostine(Ethyol) or mesna (Mesnex).

Exemplary interferons include, but are not limited to, interferonalfa-2b (Intron A) or interferon alfa-2a (Roferon-A).

Exemplary polyclonal or monoclonal antibodies include, but are notlimited to, trastuzumab (Herceptin); ofatumumab (Arzerra); bevacizumab(Avastin); rituximab (Rituxan); cetuximab (Erbitux); panitumumab(Vectibix); tositumomab/iodine131 tositumomab (Bexxar); alemtuzumab(Campath); ibritumomab (Zevalin; In-111; Y-90 Zevalin); gemtuzumab(Mylotarg); eculizumab (Soliris) ordenosumab.

Exemplary EGFR inhibitors include, but are not limited to, gefitinib(Iressa); lapatinib (Tykerb); cetuximab (Erbitux); erlotinib (Tarceva);panitumumab (Vectibix); PKI-166; canertinib (CI-1033); matuzumab(Emd7200) or EKB-569.

Exemplary HER2 inhibitors include, but are not limited to, trastuzumab(Herceptin); lapatinib (Tykerb) or AC-480.

Histone Deacetylase Inhibitors include, but are not limited to,vorinostat (Zolinza).

Exemplary hormones include, but are not limited to, tamoxifen (Soltamox;Nolvadex); raloxifene (Evista); megestrol (Megace); leuprolide (Lupron;Lupron Depot; Eligard; Viadur); fulvestrant (Faslodex); letrozole(Femara); triptorelin (Trelstar LA; Trelstar Depot); exemestane(Aromasin); goserelin (Zoladex); bicalutamide (Casodex); anastrozole(Arimidex); fluoxymesterone (Androxy; Halotestin); medroxyprogesterone(Provera; Depo-Provera); estramustine (Emcyt); flutamide (Eulexin);toremifene (Fareston); degarelix (Firmagon); nilutamide (Nilandron);abarelix (Plenaxis); or testolactone (Teslac).

Exemplary mitotic inhibitors include, but are not limited to, paclitaxel(Taxol; Onxol; Abraxane); docetaxel (Taxotere); vincristine (Oncovin;Vincasar PFS); vinblastine (Velban); etoposide (Toposar; Etopophos;VePesid); teniposide (Vumon); ixabepilone (Ixempra); nocodazole;epothilone; vinorelbine (Navelbine); camptothecin (CPT); irinotecan(Camptosar); topotecan (Hycamtin); amsacrine or lamellarin D (LAM-D).

Exemplary MTOR inhibitors include, but are not limited to, everolimus(Afinitor) or temsirolimus (Torisel); rapamune, ridaforolimus; orAP23573.

Exemplary multi-kinase inhibitors include, but are not limited to,sorafenib (Nexavar); sunitinib (Sutent); BIBW 2992; E7080; Zd6474;PKC-412; motesanib; or AP24534.

Exemplary serine/threonine kinase inhibitors include, but are notlimited to, ruboxistaurin; eril/easudil hydrochloride; flavopiridol;seliciclib (CYC202; Roscovitrine); SNS-032 (BMS-387032); Pkc412;bryostatin; KAI-9803; SF1126; VX-680; Azd1152; Arry-142886 (AZD-6244);SCIO-469; GW681323; CC-401; CEP-1347 or PD 332991.

Exemplary tyrosine kinase inhibitors include, but are not limited to,erlotinib (Tarceva); gefitinib (Iressa); imatinib (Gleevec); sorafenib(Nexavar); sunitinib (Sutent); trastuzumab (Herceptin); bevacizumab(Avastin); rituximab (Rituxan); lapatinib (Tykerb); cetuximab (Erbitux);panitumumab (Vectibix); everolimus (Afinitor); alemtuzumab (Campath);gemtuzumab (Mylotarg); temsirolimus (Torisel); pazopanib (Votrient);dasatinib (Sprycel); nilotinib (Tasigna); vatalanib (Ptk787; ZK222584);CEP-701; SU5614; MLN518; XL999; VX-322; Azd0530; BMS-354825; SKI-606CP-690; AG-490; WHI-P154; WHI-P131; AC-220; or AMG888.

Exemplary VEGF/VEGFR inhibitors include, but are not limited to,bevacizumab (Avastin); sorafenib (Nexavar); sunitinib (Sutent);ranibizumab; pegaptanib; or vandetinib.

Exemplary microtubule targeting drugs include, but are not limited to,paclitaxel, docetaxel, vincristin, vinblastin, nocodazole, epothilonesand navelbine.

Exemplary topoisomerase poison drugs include, but are not limited to,teniposide, etoposide, adriamycin, camptothecin, daunorubicin,dactinomycin, mitoxantrone, amsacrine, epirubicin and idarubicin.

Exemplary taxanes or taxane derivatives include, but are not limited to,paclitaxel and docetaxol.

Exemplary general chemotherapeutic, anti-neoplastic, anti-proliferativeagents include, but are not limited to, altretamine (Hexalen);isotretinoin (Accutane; Amnesteem; Claravis; Sotret); tretinoin(Vesanoid); azacitidine (Vidaza); bortezomib (Velcade) asparaginase(Elspar); levamisole (Ergamisol); mitotane (Lysodren); procarbazine(Matulane); pegaspargase (Oncaspar); denileukin diftitox (Ontak);porfimer (Photofrin); aldesleukin (Proleukin); lenalidomide (Revlimid);bexarotene (Targretin); thalidomide (Thalomid); temsirolimus (Torisel);arsenic trioxide (Trisenox); verteporfin (Visudyne); mimosine(Leucenol); (1M tegafur—0.4 M 5-chloro-2,4-dihydroxypyrimidine—1 Mpotassium oxonate) or lovastatin.

In another aspect, the second chemotherapeutic agent can be a cytokinesuch as G-CSF (granulocyte colony stimulating factor). In anotheraspect, a compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, analog or derivative thereof, maybe administered in combination with radiation therapy. Radiation therapycan also be administered in combination with a compound of the presentinvention and another chemotherapeutic agent described herein as part ofa multiple agent therapy. In yet another aspect, a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug,metabolite, analog or derivative thereof, may be administered incombination with standard chemotherapy combinations such as, but notrestricted to, CMF (cyclophosphamide, methotrexate and 5-fluorouracil),CAF (cyclophosphamide, adriamycin and 5-fluorouracil), AC (adriamycinand cyclophosphamide), FEC (5-fluorouracil, epirubicin, andcyclophosphamide), ACT or ATC (adriamycin, cyclophosphamide, andpaclitaxel), rituximab, Xeloda (capecitabine), Cisplatin (CDDP),Carboplatin, TS-1 (tegafur, gimestat and otastat potassium at a molarratio of 1:0.4:1), Camptothecin-11 (CPT-11, Irinotecan or Camptosar™),CHOP (cyclophosphamide, hydroxydaunorubicin, oncovin, and prednisone orprednisolone), R-CHOP (rituximab, cyclophosphamide, hydroxydaunorubicin,oncovin, prednisone or prednisolone), or CMFP (cyclophosphamide,methotrexate, 5-fluorouracil and prednisone).

In preferred embodiments, a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, may be administered with an inhibitor of an enzyme,such as a receptor or non-receptor kinase. Receptor and non-receptorkinases are, for example, tyrosine kinases or serine/threonine kinases.Kinase inhibitors described herein are small molecules, polynucleicacids, polypeptides, or antibodies.

Exemplary kinase inhibitors include, but are not limited to, Bevacizumab(targets VEGF), BIBW 2992 (targets EGFR and Erb2), Cetuximab/Erbitux(targets Erb1), Imatinib/Gleevic (targets Bcr-Abl), Trastuzumab (targetsErb2), Gefitinib/Iressa (targets EGFR), Ranibizumab (targets VEGF),Pegaptanib (targets VEGF), Erlotinib/Tarceva (targets Erb1), Nilotinib(targets Bcr-Abl), Lapatinib (targets Erb1 and Erb2/Her2),GW-572016/lapatinib ditosylate (targets HER2/Erb2), Panitumumab/Vectibix(targets EGFR), Vandetinib (targets RET/VEGFR), E7080 (multiple targetsincluding RET and VEGFR), Herceptin (targets HER2/Erb2), PKI-166(targets EGFR), Canertinib/CI-1033 (targets EGFR),Sunitinib/SU-11464/Sutent (targets EGFR and FLT3), Matuzumab/Emd7200(targets EGFR), EKB-569 (targets EGFR), Zd6474 (targets EGFR and VEGFR),PKC-412 (targets VEGR and FLT3), Vatalanib/Ptk787/ZK222584 (targetsVEGR), CEP-701 (targets FLT3), SU5614 (targets FLT3), MLN518 (targetsFLT3), XL999 (targets FLT3), VX-322 (targets FLT3), Azd0530 (targetsSRC), BMS-354825 (targets SRC), SKI-606 (targets SRC), CP-690 (targetsJAK), AG-490 (targets JAK), WHI-P154 (targets JAK), WHI-P131 (targetsJAK), sorafenib/Nexavar (targets RAF kinase, VEGFR-1, VEGFR-2, VEGFR-3,PDGFR-B, KIT, FLT-3, and RET), Dasatinib/Sprycel (BCR/ABL and Src),AC-220 (targets Flt3), AC-480 (targets all HER proteins, “panHER”),Motesanib diphosphate (targets VEGF 1-3, PDGFR, and c-kit), Denosumab(targets RANKL, inhibits SRC), AMG888 (targets HER3), and AP24534(multiple targets including Flt3).

Exemplary serine/threonine kinase inhibitors include, but are notlimited to, Rapamune (targets mTOR/FRAP1), Deforolimus (targets mTOR),Certican/Everolimus (targets mTOR/FRAP1), AP23573 (targets mTOR/FRAP1),Eril/Fasudil hydrochloride (targets RHO), Flavopiridol (targets CDK),Seliciclib/CYC202/Roscovitrine (targets CDK), SNS-032/BMS-387032(targets CDK), Ruboxistaurin (targets PKC), Pkc412 (targets PKC),Bryostatin (targets PKC), KAI-9803 (targets PKC), SF 1126 (targetsPI3K), VX-680 (targets Aurora kinase), Azdl 152 (targets Aurora kinase),Arry-142886/AZD-6244 (targets MAP/MEK), SCIO-469 (targets MAP/MEK),GW681323 (targets MAP/MEK), CC-401 (targets JNK), CEP-1347 (targetsJNK), and PD 332991 (targets CDK).

The disorder in which EZH2-mediated protein methylation plays a part canbe a neurological disease. The compounds of this invention can thus alsobe used for treating or studying neurologic diseases such as epilepsy,schizophrenia, bipolar disorder or other psychological and/orpsychiatric disorders, neuropathies, skeletal muscle atrophy, andneurodegenerative diseases, e.g., a neurodegenerative disease. Exemplaryneurodegenerative diseases include: Alzheimer's, Amyotrophic LateralSclerosis (ALS), and Parkinson's disease. Another class ofneurodegenerative diseases includes diseases caused at least in part byaggregation of poly-glutamine. Diseases of this class include:Huntington's Diseases, Spinalbulbar Muscular Atrophy (SBMA or Kennedy'sDisease) Dentatorubropallidoluysian Atrophy (DRPLA), SpinocerebellarAtaxia 1 (SCA1), Spinocerebellar Ataxia 2 (SCA2), Machado-Joseph Disease(MJD; SCA3), Spinocerebellar Ataxia 6 (SCA6), Spinocerebellar Ataxia 7(SCA7), and Spinocerebellar Ataxia 12 (SCA12).

Any other disease in which epigenetic methylation, which is mediated byEZH2, plays a role may be treatable or preventable using compounds andmethods described herein, or such diseases and potential treatmentsthereof may be studied with the compounds described herein.

4. PHARMACEUTICAL COMPOSITIONS

The present invention also provides pharmaceutical compositionscomprising a compound of any Formula disclosed herein in combinationwith at least one pharmaceutically acceptable excipient or carrier.

A “pharmaceutical composition” is a formulation containing the compoundsof the present invention in a form suitable for administration to asubject. In one embodiment, the pharmaceutical composition is in bulk orin unit dosage form. The unit dosage form is any of a variety of forms,including, for example, a capsule, an IV bag, a tablet, a single pump onan aerosol inhaler or a vial. The quantity of active ingredient (e.g., aformulation of the disclosed compound or salt, hydrate, solvate orisomer thereof) in a unit dose of composition is an effective amount andis varied according to the particular treatment involved. One skilled inthe art will appreciate that it is sometimes necessary to make routinevariations to the dosage depending on the age and condition of thepatient. The dosage will also depend on the route of administration. Avariety of routes are contemplated, including oral, pulmonary, rectal,parenteral, transdermal, subcutaneous, intravenous, intramuscular,intraperitoneal, inhalational, buccal, sublingual, intrapleural,intrathecal, intranasal, and the like. Dosage forms for the topical ortransdermal administration of a compound of this invention includepowders, sprays, ointments, pastes, creams, lotions, gels, solutions,patches and inhalants. In one embodiment, the active compound is mixedunder sterile conditions with a pharmaceutically acceptable carrier, andwith any preservatives, buffers, or propellants that are required.

As used herein, the phrase “pharmaceutically acceptable” refers to thosecompounds, anions, cations, materials, compositions, carriers, and/ordosage forms which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of human beings and animalswithout excessive toxicity, irritation, allergic response, or otherproblem or complication, commensurate with a reasonable benefit/riskratio.

“Pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic and neither biologically nor otherwise undesirable, andincludes excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (topical), andtransmucosal administration. Solutions or suspensions used forparenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid;buffers such as acetates, citrates or phosphates, and agents for theadjustment of tonicity such as sodium chloride or dextrose. The pH canbe adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

A compound or pharmaceutical composition of the invention can beadministered to a subject in many of the well-known methods currentlyused for chemotherapeutic treatment. For example, for treatment ofcancers, a compound of the invention may be injected directly intotumors, injected into the blood stream or body cavities or taken orallyor applied through the skin with patches. The dose chosen should besufficient to constitute effective treatment but not so high as to causeunacceptable side effects. The state of the disease condition (e.g.,cancer, precancer, and the like) and the health of the patient shouldpreferably be closely monitored during and for a reasonable period aftertreatment.

The term “therapeutically effective amount”, as used herein, refers toan amount of a pharmaceutical agent to treat, ameliorate, or prevent anidentified disease or condition, or to exhibit a detectable therapeuticor inhibitory effect. The effect can be detected by any assay methodknown in the art. The precise effective amount for a subject will dependupon the subject's body weight, size, and health; the nature and extentof the condition; and the therapeutic or combination of therapeuticsselected for administration. Therapeutically effective amounts for agiven situation can be determined by routine experimentation that iswithin the skill and judgment of the clinician. In a preferred aspect,the disease or condition to be treated is cancer. In another aspect, thedisease or condition to be treated is a cell proliferative disorder.

For any compound, the therapeutically effective amount can be estimatedinitially either in cell culture assays, e.g., of neoplastic cells, orin animal models, usually rats, mice, rabbits, dogs, or pigs. The animalmodel may also be used to determine the appropriate concentration rangeand route of administration. Such information can then be used todetermine useful doses and routes for administration in humans.Therapeutic/prophylactic efficacy and toxicity may be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., ED₅₀ (the dose therapeutically effective in 50% of thepopulation) and LD₅₀ (the dose lethal to 50% of the population). Thedose ratio between toxic and therapeutic effects is the therapeuticindex, and it can be expressed as the ratio, LD₅₀/ED₅₀. Pharmaceuticalcompositions that exhibit large therapeutic indices are preferred. Thedosage may vary within this range depending upon the dosage formemployed, sensitivity of the patient, and the route of administration.

Dosage and administration are adjusted to provide sufficient levels ofthe active agent(s) or to maintain the desired effect. Factors which maybe taken into account include the severity of the disease state, generalhealth of the subject, age, weight, and gender of the subject, diet,time and frequency of administration, drug combination(s), reactionsensitivities, and tolerance/response to therapy. Long-actingpharmaceutical compositions may be administered every 3 to 4 days, everyweek, or once every two weeks depending on half-life and clearance rateof the particular formulation.

The pharmaceutical compositions containing active compounds of thepresent invention may be manufactured in a manner that is generallyknown, e.g., by means of conventional mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping, orlyophilizing processes. Pharmaceutical compositions may be formulated ina conventional manner using one or more pharmaceutically acceptablecarriers comprising excipients and/or auxiliaries that facilitateprocessing of the active compounds into preparations that can be usedpharmaceutically. Of course, the appropriate formulation is dependentupon the route of administration chosen.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringeability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), and suitable mixturesthereof. The proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as manitol and sorbitol, and sodium chloridein the composition. Prolonged absorption of the injectable compositionscan be brought about by including in the composition an agent whichdelays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound in the required amount in an appropriate solvent with one or acombination of ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the active compound into a sterile vehicle that contains abasic dispersion medium and the required other ingredients from thoseenumerated above. In the case of sterile powders for the preparation ofsterile injectable solutions, methods of preparation are vacuum dryingand freeze-drying that yields a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

Oral compositions generally include an inert diluent or an ediblepharmaceutically acceptable carrier. They can be enclosed in gelatincapsules or compressed into tablets. For the purpose of oral therapeuticadministration, the active compound can be incorporated with excipientsand used in the form of tablets, troches, or capsules. Oral compositionscan also be prepared using a fluid carrier for use as a mouthwash,wherein the compound in the fluid carrier is applied orally and swishedand expectorated or swallowed. Pharmaceutically compatible bindingagents, and/or adjuvant materials can be included as part of thecomposition. The tablets, pills, capsules, troches and the like cancontain any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate or Sterotes; a glidant such as colloidal silicondioxide; a sweetening agent such as sucrose or saccharin; or a flavoringagent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser, whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, detergents, bile salts, andfusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

The active compounds can be prepared with pharmaceutically acceptablecarriers that will protect the compound against rapid elimination fromthe body, such as a controlled release formulation, including implantsand microencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved.

In therapeutic applications, the dosages of the pharmaceuticalcompositions used in accordance with the invention vary depending on theagent, the age, weight, and clinical condition of the recipient patient,and the experience and judgment of the clinician or practitioneradministering the therapy, among other factors affecting the selecteddosage. Generally, the dose should be sufficient to result in slowing,and preferably regressing, the growth of the tumors and also preferablycausing complete regression of the cancer. Dosages can range from about0.01 mg/kg per day to about 5000 mg/kg per day. In preferred aspects,dosages can range from about 1 mg/kg per day to about 1000 mg/kg perday. In an aspect, the dose will be in the range of about 0.1 mg/day toabout 50 g/day; about 0.1 mg/day to about 25 g/day; about 0.1 mg/day toabout 10 g/day; about 0.1 mg to about 3 g/day; or about 0.1 mg to about1 g/day, in single, divided, or continuous doses (which dose may beadjusted for the patient's weight in kg, body surface area in m², andage in years). An effective amount of a pharmaceutical agent is thatwhich provides an objectively identifiable improvement as noted by theclinician or other qualified observer. For example, regression of atumor in a patient may be measured with reference to the diameter of atumor. Decrease in the diameter of a tumor indicates regression.Regression is also indicated by failure of tumors to reoccur aftertreatment has stopped. As used herein, the term “dosage effectivemanner” refers to amount of an active compound to produce the desiredbiological effect in a subject or cell.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

The compounds of the present invention are capable of further formingsalts. All of these forms are also contemplated within the scope of theclaimed invention.

As used herein, “pharmaceutically acceptable salts” refer to derivativesof the compounds of the present invention wherein the parent compound ismodified by making acid or base salts thereof. Examples ofpharmaceutically acceptable salts include, but are not limited to,mineral or organic acid salts of basic residues such as amines, alkalior organic salts of acidic residues such as carboxylic acids, and thelike. The pharmaceutically acceptable salts include the conventionalnon-toxic salts or the quaternary ammonium salts of the parent compoundformed, for example, from non-toxic inorganic or organic acids. Forexample, such conventional non-toxic salts include, but are not limitedto, those derived from inorganic and organic acids selected from2-acetoxybenzoic, 2-hydroxyethane sulfonic, acetic, ascorbic, benzenesulfonic, benzoic, bicarbonic, carbonic, citric, edetic, ethanedisulfonic, 1,2-ethane sulfonic, fumaric, glucoheptonic, gluconic,glutamic, glycolic, glycollyarsanilic, hexylresorcinic, hydrabamic,hydrobromic, hydrochloric, hydroiodic, hydroxymaleic, hydroxynaphthoic,isethionic, lactic, lactobionic, lauryl sulfonic, maleic, malic,mandelic, methane sulfonic, napsylic, nitric, oxalic, pamoic,pantothenic, phenylacetic, phosphoric, polygalacturonic, propionic,salicyclic, stearic, subacetic, succinic, sulfamic, sulfanilic,sulfuric, tannic, tartaric, toluene sulfonic, and the commonly occurringamine acids, e.g., glycine, alanine, phenylalanine, arginine, etc.

Other examples of pharmaceutically acceptable salts include hexanoicacid, cyclopentane propionic acid, pyruvic acid, malonic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonicacid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid,camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylicacid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylaceticacid, muconic acid, and the like. The present invention also encompassessalts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like. In the salt form, it is understood thatthe ratio of the compound to the cation or anion of the salt can be 1:1,or any ration other than 1:1, e.g., 3:1, 2:1, 1:2, or 1:3.

It should be understood that all references to pharmaceuticallyacceptable salts include solvent addition forms (solvates) or crystalforms (polymorphs) as defined herein, of the same salt.

The compounds of the present invention can also be prepared as esters,for example, pharmaceutically acceptable esters. For example, acarboxylic acid function group in a compound can be converted to itscorresponding ester, e.g., a methyl, ethyl or other ester. Also, analcohol group in a compound can be converted to its corresponding ester,e.g., acetate, propionate or other ester.

The compounds of the present invention can also be prepared as prodrugs,for example, pharmaceutically acceptable prodrugs. The terms “pro-drug”and “prodrug” are used interchangeably herein and refer to any compoundwhich releases an active parent drug in vivo. Since prodrugs are knownto enhance numerous desirable qualities of pharmaceuticals (e.g.,solubility, bioavailability, manufacturing, etc.), the compounds of thepresent invention can be delivered in prodrug form. Thus, the presentinvention is intended to cover prodrugs of the presently claimedcompounds, methods of delivering the same and compositions containingthe same. “Prodrugs” are intended to include any covalently bondedcarriers that release an active parent drug of the present invention invivo when such prodrug is administered to a subject. Prodrugs in thepresent invention are prepared by modifying functional groups present inthe compound in such a way that the modifications are cleaved, either inroutine manipulation or in vivo, to the parent compound. Prodrugsinclude compounds of the present invention wherein a hydroxy, amino,sulfhydryl, carboxy or carbonyl group is bonded to any group that may becleaved in vivo to form a free hydroxyl, free amino, free sulfhydryl,free carboxy or free carbonyl group, respectively.

Examples of prodrugs include, but are not limited to, esters (e.g.,acetate, dialkylaminoacetates, formates, phosphates, sulfates andbenzoate derivatives) and carbamates (e.g., N,N-dimethylaminocarbonyl)of hydroxy functional groups, esters (e.g., ethyl esters,morpholinoethanol esters) of carboxyl functional groups, N-acylderivatives (e.g., N-acetyl)N-Mannich bases, Schiff bases and enaminonesof amino functional groups, oximes, acetals, ketals and enol esters ofketone and aldehyde functional groups in compounds of the invention, andthe like, See Bundegaard, H., Design of Prodrugs, p1-92, Elesevier, NewYork-Oxford (1985).

The compounds, or pharmaceutically acceptable salts, esters or prodrugsthereof, are administered orally, nasally, transdermally, pulmonary,inhalationally, buccally, sublingually, intraperintoneally,subcutaneously, intramuscularly, intravenously, rectally,intrapleurally, intrathecally and parenterally. In one embodiment, thecompound is administered orally. One skilled in the art will recognizethe advantages of certain routes of administration.

The dosage regimen utilizing the compounds is selected in accordancewith a variety of factors including type, species, age, weight, sex andmedical condition of the patient; the severity of the condition to betreated; the route of administration; the renal and hepatic function ofthe patient; and the particular compound or salt thereof employed. Anordinarily skilled physician or veterinarian can readily determine andprescribe the effective amount of the drug required to prevent, counter,or arrest the progress of the condition.

Techniques for formulation and administration of the disclosed compoundsof the invention can be found in Remington: the Science and Practice ofPharmacy, 19^(th) edition, Mack Publishing Co., Easton, Pa. (1995). Inan embodiment, the compounds described herein, and the pharmaceuticallyacceptable salts thereof, are used in pharmaceutical preparations incombination with a pharmaceutically acceptable carrier or diluent.Suitable pharmaceutically acceptable carriers include inert solidfillers or diluents and sterile aqueous or organic solutions. Thecompounds will be present in such pharmaceutical compositions in amountssufficient to provide the desired dosage amount in the range describedherein.

All percentages and ratios used herein, unless otherwise indicated, areby weight. Other features and advantages of the present invention areapparent from the different examples. The provided examples illustratedifferent components and methodology useful in practicing the presentinvention. The examples do not limit the claimed invention. Based on thepresent disclosure the skilled artisan can identify and employ othercomponents and methodology useful for practicing the present invention.

In the synthetic schemes described herein, compounds may be drawn withone particular configuration for simplicity. Such particularconfigurations are not to be construed as limiting the invention to oneor another isomer, tautomer, regioisomer or stereoisomer, nor does itexclude mixtures of isomers, tautomers, regioisomers or stereoisomers;however, it will be understood that a given isomer, tautomer,regioisomer or stereoisomer may have a higher level of activity thananother isomer, tautomer, regioisomer or stereoisomer.

Compounds designed, selected and/or optimized by methods describedabove, once produced, can be characterized using a variety of assaysknown to those skilled in the art to determine whether the compoundshave biological activity. For example, the molecules can becharacterized by conventional assays, including but not limited to thoseassays described below, to determine whether they have a predictedactivity, binding activity and/or binding specificity.

Furthermore, high-throughput screening can be used to speed up analysisusing such assays. As a result, it can be possible to rapidly screen themolecules described herein for activity, using techniques known in theart. General methodologies for performing high-throughput screening aredescribed, for example, in Devlin (1998) High Throughput Screening,Marcel Dekker; and U.S. Pat. No. 5,763,263. High-throughput assays canuse one or more different assay techniques including, but not limitedto, those described below.

All publications and patent documents cited herein are incorporatedherein by reference as if each such publication or document wasspecifically and individually indicated to be incorporated herein byreference. Citation of publications and patent documents is not intendedas an admission that any is pertinent prior art, nor does it constituteany admission as to the contents or date of the same. The inventionhaving now been described by way of written description, those of skillin the art will recognize that the invention can be practiced in avariety of embodiments and that the foregoing description and examplesbelow are for purposes of illustration and not limitation of the claimsthat follow.

5. EXAMPLES General Experimental NMR

¹H-NMR spectra were taken using CDCl₃ unless otherwise stated and wererecorded at 400 or 500 MHz using a Varian or Oxford instruments magnet(500 MHz) instruments. Multiplicities indicated are s=singlet,d=doublet, t=triplet, q=quartet, quint=quintet, sxt=sextet, m=multiplet,dd=doublet of doublets, dt=doublet of triplets; br indicates a broadsignal.

LCMS and HPLC

Shimadzu LC-Q, Shimadzu LCMS-2010EV or Waters Acquity Ultra PerformanceLC. HPLC: Products were analyzed by Shimadzu SPD-20A with 150×4.5 mm YMCODS-M80 column or 150×4.6 mm YMC-Pack Pro C18 column at 1.0 ml/min.

Mobile phase was MeCN:H2O=3:2 (containing 0.3% SDS and 0.05% H₃PO₄),

0.05% TFA in water, 0.05% TFA in acetonitrile (gradient Initial 20%,then 0.05% TFA/MeCN to conc. to 95% in 3 min. holds for 0.5 min. at 3.51to 4.50 min then 0.05% TFA/MeCN conc. 20%).

Alternatively the LCMS, 2 different methods were used; the one we usethe most is the high pH (METCR1600) and the other one for more standardcompounds (METCR1416).

0.1% Formic acid in water—Mobile phase “A” 0.1% Formic acid inacetonitrile—Mobile phase “B” utilizing Waters Atlantis dC18, 2.1 mm×100mm, 3 μm column, with a flow rate=0.6 ml/min Column temperature=40° C.;Time (mins) % B 0.00 min 5% B. 5.0 mins 100% B, 5.4 mins 100% B and 0.42mins 5% B

3.5 minute method refers to Atlantis dC18, 2.1 mm×50 mm, 3 m column,flow rate of 1 ml/min at 40 C. Mobile phase A Formic acid (aq.) 0.1%mobile phase B formic acid (MeCN) 0.1%, injection 3 μL, gradient 0 mins(5% organic), 2.5 min (100% organic), 2.7 mins (100% organic), 2.71 min(5% organic), 3.5 min (5% organic)

7.0 minute method refers to Atlantis dC18, 2.1 mm×100 mm, 3 μm column,flow rate of 0.6 ml/min at 40 C. Mobile phase A Formic acid (aq.) 0.1%mobile phase B formic acid (MeCN) 0.1%, injection 3 μL, gradient 0 mins(5% organic), 5 min (100% organic), 5.4 mins (100% organic), 5.42 min(5% organic), 7 min (5% organic)

Both the 3. 5 and 7 minute methods were performed on a MS18 ShimadzuLCMS-2010EV or a MS19 Shimadzu LCMS-2010EV system utilizing LC-20ABpumps and SPD-M20A PDA detectors.

Products were purified by HPLC/MS using Waters AutoPurification Systemwith 3100 Mass Detector.

HPLC analyses may also be performed on a Shimdazu LC-2010 CHT using anYMC ODS-A, C18, (150×4.6×5 m) column at ambient temperature with a flowRate of 1.4 ml/min. An injection volume of 10 μl is utilized anddetection occurs via UV/PDA. Mobile Phase A is 0.05% TFA in water andMobile Phase B is 0.05% TFA in acetonitrile with a gradient program ofInitial 5% B to 95% B in 8 min, hold for 1.5 min, at 9.51 to 12 min B.conc. 0.5%. The diluent is the mobile phase

Other

Automated flash column chromatography was performed on a Biotage Isoleraversion 4. 10 g SNAP cartridge running at 12 ml/min or a 25 g SNAPcartridge running at 25 ml/min and detecting at 254 nm and 280 nm.

Select Nitrile reductions may be performed on a ThalesNano H-Cube®according to the conditions described in the experimental procedure.

Example 1: Synthesis of Compound 1:5-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropuridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid

To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g, 276.2 mmol)in conc. H₂SO₄ (200 mL), 1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione(43.4 g, 151.8 mmol) was added portion wise at room temperature andreaction mass was stirred at room temperature for 5 h. On completion,reaction mass was poured on ice cold water, solid precipitated wasfiltered, resulting residue was washed with water and dried under vacuumto give the desired compound (71.7 g, 100%).

Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate

To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid (287 g,1103 mmol) in DMF (150 mL), sodium carbonate (468 g, 4415 mmol) andmethyl iodide (626.63 g, 4415 mmol) were added. Resulting reaction masswas heated at 60° C. for 8 h. On completion, solid precipitated wasfiltered, residue washed with diethyl ether (5 times). Combined organiclayers were dried, concentrated under reduced pressure giving thedesired crude compound (302 g, 99%).

Step 3: methyl 3-amino-5-bromo-2-methylbenzoate

To a stirred solution of methyl 5-bromo-2-methyl-3-nitrobenzoate (150 g,544 mmol) in ethanol (750 mL), ammonium chloride (150 g, 2777 mmol)dissolved in water (750 mL) and iron powder (93.3 g, 1636 mmol) wereadded under stirring. Resulting reaction mass was heated at 80° C. for 7h. On completion, reaction mass was filtered through celite givingwashing of water and ethyl acetate, filtrate was extracted with ethylacetate. Combined organic layers were dried, concentrated under reducedpressure giving the desired compound.

Step 4: methyl 5-bromo-3-(cyclopentylamino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (0.3 g,1.33 mmol) and cyclopentanone (0.56 g, 6.6 mmol) in methanol (3 mL),acetic acid (0.159 g, 2.6 mmol) was added and reaction stirred at roomtemperature for 3 h. Then sodium cyanoborohydride (0.208 g, 3.3 mmol)was added and reaction stirred overnight. On completion, solvent wasremoved under reduced pressure to give the desired compound.

Step 5: methyl 5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate

To a stirred solution of the crude methyl5-bromo-3-(cyclopentylamino)-2-methylbenzoate (0.7 g, 2.25 mmol) inacetonitrile (15 mL), cesium carbonate (1.47 g, 4.50 mmol) and methyliodide (1.6 g, 11.26 mmol) were added; resulting reaction mass washeated at 80° C. for 7 h. On completion, reaction mass was cooled toroom temperature and filtered, residue was washed with ethyl acetate andfiltrate was concentrated and then purified by column chromatography toafford the desired compound (0.6 g, 82%).

Step 6:5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

Aqueous NaOH (0.11 g, 2.75 mmol) was added to a solution of methyl5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate (0.6 g, 1.8 mmol)in MeOH (1.5 mL) and stirred at 60° C. for 1 h. After completion of thereaction, ethanol was removed under reduced pressure and acidified usingdilute HCl up to pH 6 and pH 4 was adjusted using citric acid.Extraction was carried out using ethyl acetate. The combined organiclayers were dried and concentrated to give the respective acid (0.5 g,87%).

The acid (0.5 g, 1.60 mmol) was then dissolved in DMSO (3 mL) and3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.49 g, 3.22 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (1.25 g, 2.41 mmol) was added to it and stirring wascontinued for overnight. After completion of the reaction, reaction masswas poured into ice to obtain solid, this was filtered and washed withacetonitrile followed by ether to provide the desired compound (0.315 g,44%).

Step 7: Synthesis of5-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide

A solution of5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.), (4-formylphenyl)boronic acid (1.2 equiv.) and Pd (PPh₃)₄(0.1 equiv.) in 1,4-dioxane (4 mL) was purged with argon for 10 min.Then, a 2 M Na₂CO₃ solution (3.6 equiv.) was added to it and argon waspurged again for 10 min. The reaction mixture was stirred at 100° C. for2 h. After completion of the reaction, water was added to it andextraction was carried out using DCM. The combined organic layers werewashed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford the desired compound (0.1 g, 44%).

Step 8: Synthesis of5-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropuridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide(0.1 g, 0.212 mmol) and N,N-dimethylamine (0.047 g, 1.06 mmol) inmethanol (3 mL), acetic acid (0.1 g, 0.21 mmol) was added and reactionstirred at room temperature for 3 h. Then sodium cyanoborohydride (0.033g, 0.53 mmol) was added and reaction stirred overnight. On completion,solvent was removed under reduced pressure and residue purified bycolumn chromatography over silica gel to give the desired compound (0.04g, 37%). LCMS: 501.39 (M+1)₊; HPLC: 90.78% (@ 254 nm) (R_(t); 4.171;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H),8.17 (t, 1H), 7.57 (d, 2H, J=8 Hz), 7.33-7.37 (m, 3H), 7.17 (s, 1H),5.85 (s, 1H), 4.27 (d, 2H, J=4.4 Hz), 3.52 (t, 1H, J=7.2 Hz), 3.04 (s,2H), 2.54 (s, 3H), 2.23 (s, 3H), 2.19 (s, 3H), 2.15 (s, 6H), 2.09 (s,3H), 1.70-1.72 (m, 2H), 1.61 (m, 2H), 1.43-1.50 (m, 4H).

Example 2: Synthesis of Compound 2:5-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A solution of5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.), (4-(morpholinomethyl)phenyl)boronic acid (1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane (4 mL) was purged with argon for 10min. Then, a 2 M Na₂CO₃ solution (3.6 equiv.) was added to it and argonwas purged again for 10 min. The reaction mixture was stirred at 100° C.for 2 h. After completion of the reaction, water was added to it andextraction was carried out using DCM. The combined organic layers werewashed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford the desired compound (0.02 g, 16%). LCMS: 543.22 (M+1)⁺; HPLC:99.53% (@ 254 nm) (R_(t); 4.181; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient:5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.17 (t, 1H, J=4.4 Hz), 7.98 (s,1H), 7.73 (d, 1H, J=7.6 Hz), 7.57 (d, 2H, J=7.6 Hz), 7.37 (s, 2H), 7.17(s, 1H), 5.85 (s, 1H), 4.27 (d, 2H, J=4.8 Hz), 3.44-3.57 (m, 7H), 2.54(s, 3H), 2.32-2.36 (m, 4H), 2.23 (s, 3H), 2.19 (s, 3H), 2.09 (s, 3H),1.69-1.72 (m, 2H), 1.61 (m, 2H), 1.43-1.50 (m, 4H).

Example 3: Synthesis of5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

Aqueous NaOH (0.1 g, 2.5 mmol) was added to a solution of methyl5-bromo-3-(cyclopentylamino)-2-methylbenzoate (0.39 g, 1.25 mmol) inMeOH (5 mL) and stirred at 60° C. for 1 h. Ethanol was removed underreduced pressure, and the solution acidified using dilute HCl to pH 6and citric acid to pH 4. The product was extracted with ethyl acetateand the combined organic layers were concentrated to give the desiredacid (0.26 g, 0.82 mmol). The acid was dissolved in DMSO (3 mL) and3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.25 g, 1.68 mmol) wasadded to the solution. The reaction mixture was stirred at roomtemperature for 15 min before PYBOP (0.65 g, 1.26 mmol) was added to itand stirring was continued overnight. The reaction mixture was pouredonto ice to obtain a solid, and this solid was collected by filtrationand washed with acetonitrile followed by ether to provide5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.178 g, 50%).

Step 2: Synthesis of5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide

A solution of5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.), (4-formylphenyl)boronic acid (1.2 equiv.) and Pd (PPh₃)₄(0.1 equiv.) in 1,4-dioxane (4 mL) was purged with argon for 10 min.Then, 2 M Na₂CO₃ solution (3.6 equiv.) was added to it and argon waspurged again for 10 min. The reaction mixture was stirred at 100° C. for2 h. After cooling to room temperature water was added to the mixtureand then product was extracted with DCM. The combined organic layerswere washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide.

Step 3:5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide(0.11 g, 0.24 mmol) and N,N-dimethylamine (0.044 g, 1.2 mmol) inmethanol (3 mL) was added acetic acid (0.014 g, 0.24 mmol) and thesolution stirred at room temperature for 3 h. Then sodiumcyanoborohydride (0.030 g, 0.48 mmol) was added and the solution stirredovernight. The solvent was removed under reduced pressure and theresidue purified by column chromatography over silica gel to afforddesired5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

LCMS: 486.21 (M+1)⁺; HPLC: 99.84% (@ 254 nm) (R_(t); 4.799; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.44 (s, 1H),8.02-8.03 (m, 1H), 7.62 (d, 2H, J=7.6 Hz), 7.44 (s, 2H), 6.80 (s, 1H),6.73 (s, 1H), 5.85 (s, 1H), 4.65 (d, 1H, J=6.4 Hz), 4.27 (d, 2H, J=4.4Hz), 3.89 (d, 2H, J=5.2 Hz), 2.49 (7H merged in Solvent Peak), 1.98-2.19(m, 11H), 1.55-1.70 (m, 6H).

Example 4: Synthesis of5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A solution of5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.), (4-(morpholinomethyl)phenyl)boronic acid (1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane (4 mL) was purged with argon for 10min. Then, 2 M Na₂CO₃ solution (3.6 equiv.) was added to it and argonwas purged again for 10 min. The reaction mixture was stirred at 100° C.for 2 h. After completion of the reaction, water was added to it andextraction was carried out using DCM. The combined organic layers werewashed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford5-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamidewhich was further purified using preparative HPLC which gave the TFAsalt.

LCMS: 529.30 (M+1)+; HPLC: 99.46% (@ 254 nm) (Rt; 4.782; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); 1H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 9.90 (s, 1H), 8.06 (s,1H), 7.72 (d, 2H, J=8 Hz), 7.55 (d, 2H, J=8 Hz), 6.83 (s, 1H), 6.76 (s,1H), 5.86 (s, 1H), 4.37 (s, 2H), 4.27 (d, 2H, J=4 Hz), 3.89-3.98 (m,3H), 3.28-3.31 (m, 2H), 3.14 (s, 2H), 2.19 (s, 3H), 2.10 (s, 3H), 2.05(s, 3H), 1.98-1.99 (m, 2H), 1.70 (s, 2H), 1.55 (s, 4H).

Example 5: Synthesis of2-(cyclohex-1-en-1-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)isonicotinamide

Step 1: Synthesis of methyl2-chloro-6-(4-(hydroxymethyl)phenyl)isonicotinate

A solution of methyl 2,6-dichloroisonicotinate (1 g, 4.85 mmol), boronicacid (0.73 g, 4.8 mmol) and PdCl₂(PPh₃)₂ (0.15 g, 0.218 mmol) in THF (20mL) was degassed for 15 min. Then Cs₂CO₃ was added and reaction masspurged again for 10 min. Reaction was heated at 70° C. for 2 h. Oncompletion, reaction mass was concentrated and purified by columnchromatography over silica gel affording methyl2-chloro-6-(4-(hydroxymethyl)phenyl)isonicotinate (0.45 g, 33%).

Step 2: Synthesis of methyl2-(4-(bromomethyl)phenyl)-6-chloroisonicotinate

To a solution of methyl2-chloro-6-(4-(hydroxymethyl)phenyl)isonicotinate (0.67 g, 2.418 mmol)in DCM (10 mL), triphenyl phosphine (1 g, 3.86 mmol) and carbontetrabromide (1.63 g, 3.87 mmol) were added at 0° C. and reaction massstirred for overnight at rt. On completion, reaction mass wasconcentrated and purified by column chromatography over silica gelaffording methyl 2-(4-(bromomethyl)phenyl)-6-chloroisonicotinate (0.53g, 64%).

Step 3: Synthesis of methyl2-chloro-6-(4-((dimethylamino)methyl)phenyl)isonicotinate

To a solution of methyl 2-(4-(bromomethyl)phenyl)-6-chloroisonicotinate(0.533 g, 1.56 mmol) in THF, dimethylamine (7.8 mL, 2M solution in THF)was added and reaction mass stirred at rt for overnight. On completion,reaction mass concentrated and crude obtained was purified by columnchromatography over silica gel obtaining pure methyl2-chloro-6-(4-((dimethylamino)methyl)phenyl)isonicotinate (0.48 g, 99%).

Step 4: Synthesis of2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)isonicotinamide

To a solution of methyl2-chloro-6-(4-((dimethylamino)methyl)phenyl)isonicotinate (0.48 g, 1.578mmol) in ethanol (5 mL), NaOH (0.094 g, 2.368 mmol), dissolved in water(1 mL), was added and reaction mass heated at 60° C. for 1 h. Oncompletion, solvent was evaporated under reduced pressure. Residue waswashed with ether and acidified with 1N HCl till pH 8 and then withcitric acid till pH 5-6. Aqueous layer was extracted with 20% MeOH/DCMand combined organic layers were concentrated under reduced pressure toafford the acid (0.47 g, crude) which was used in next step withoutfurther purification. To a solution of this acid (0.47 g, 1.64 mmol) inDMSO (4 mL), PyBOP (1.26 g, 2.43 mmol) was added and reaction stirred atrt for 15 min. Then 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.49g, 3.28 mmol) was added and reaction stirred overnight. On completion,water was added and aqueous layer extracted with 20% MeOH/DCM. Combinedorganic layers were concentrated and residue purified by silica gelcolumn chromatography affording2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)isonicotinamide(0.3 g, 43.6%)

Step 5: Synthesis of2-(cyclohex-1-en-1-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)isonicotinamide

To a stirred solution of2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)isonicotinamide(0.11 g, 0.25 mmol), boronic acid (0.059 g, 0.27 mmol) in dioxane/watermixture (3 mL+1.5 mL), Na₂CO₃ (0.098 g, 3.6 mmol) was added and reactionmass purged with argon for 15 min. Then Pd(PPh₃)₄ (0.028 g, 0.025 mmol)was added and argon was purged again for 10 min. Reaction mass washeated at 100° C. for 3 h. On completion, reaction mass filtered throughcelite and celite bed washed with ethyl acetate. Combined filtrates weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column over silica gel to obtain2-(cyclohex-1-en-1-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)isonicotinamide.

Analytical Data: LCMS: 471.30 (M+1)⁺; HPLC: 95.64% (@ 254 nm) (R_(t);5.661; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52(s, 1H), 8.79 (t, 1H), 8.13 (s, 1H), 8.10 (d, 2H, J=7.60 Hz), 7.81 (s,1H), 7.41 (d, 2H, J=7.60 Hz), 6.90 (bs, 1H), 5.88 (s, 1H), 4.34 (d, 2H,J=4.8 Hz), 3.44 (s, 2H), 2.56 (bs, 2H), 2.26 (bs, 2H), 2.18 (s, 3H),2.17 (s, 6H), 2.12 (s, 3H), 1.80-1.72 (m, 2H), 1.68-1.60 (m, 2H).

Example 6: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-((dimethylamino)methyl)phenyl)-6-(piperidin-1-yl)isonicotinamide

Step 1: Synthesis of methyl 2-chloro-6-(piperidin-1-yl)isonicotinate

A solution of methyl 2,6-dichloroisonicotinate (1 g, 4.85 mmol),piperidine (0.61 g, 7.28 mmol), K₂CO₃ (1.38 g, 9.7 mmol) in acetonitrile(20 mL) was heated at 90° C. for 20 h. After completion of reaction,reaction mass was filtered, filtrate concentrated and purified by columnto obtain pure methyl 2-chloro-6-(piperidin-1-yl)isonicotinate (1.23 g,90%).

Step 2: Synthesis of2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(piperidin-1-yl)isonicotinamide

To a solution of methyl 2-chloro-6-(piperidin-1-yl)isonicotinate (1.1 g,4.33 mmol) in ethanol (10 mL), NaOH (0.207 g, 5.196 mmol), dissolved inwater (2 mL), was added and reaction mass heated at 60° C. for 1 h. Oncompletion, solvent was evaporated under reduced pressure. Residue waswashed with ether and acidified with 1N HCl till pH 8 and then withcitric acid till pH 5-6. Solid obtained was filtered, washed with waterand finally dried under reduced pressure to afford the acid (0.92 g,89%) which was used in next step without further purification. To asolution of this acid (0.9 g, 3.75 mmol) in DMSO (10 mL), PyBOP (3.9 g,7.5 mmol) was added and reaction stirred at rt for 15 min. Then3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (1.5 g, 10 mmol) was addedand reaction stirred overnight. On completion, water was added and solidthat precipitates out was filter, washed with water and dried to obtain2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(piperidin-1-yl)isonicotinamide(1 g, 74%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-formylphenyl)-6-(piperidin-1-yl)isonicotinamide

To a stirred solution of2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(piperidin-1-yl)isonicotinamide(0.6 g, 1.6 mmol), boronic acid (0.263 g, 1.76 mmol) in dioxane/watermixture (15 mL+5 mL), Na₂CO₃ (0.61 g, 5.76 mmol) was added and reactionmass purged for 15 min with argon. Then Pd(PPh₃)₄ (0.184 g, 0.16 mmol)was added and argon was purged again for 10 min. Reaction mass washeated at 100° C. for 3 h. On completion, reaction mass filtered throughcelite and celite bed washed with ethyl acetate. Combined filtrates weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column over silica gel to obtainN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-formylphenyl)-6-(piperidin-1-yl)isonicotinamide(0.5 g, 71%).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-((dimethylamino)methyl)phenyl)-6-(piperidin-1-yl)isonicotinamide

To a solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-formylphenyl)-6-(piperidin-1-yl)isonicotinamide(0.2 g, 0.45 mmol) in methanol (12 mL), dimethyl amine (2.6 mL, 4.5mmol, 2M solution in THF) and acetic acid (0.02 g, 0.45 mmol) were addedand reaction mass stirred at rt for 90 min. Then reaction mass wascooled to 0° C. and sodium cyanoborohydride (0.056 g, 0.9 mmol) wasadded. Reaction stirred at 0° C. for 2 h and then stirred at rt forovernight. On completion, solvent was removed under reduced pressure,residue treated with water and extracted with ethyl acetate. Combinedethyl acetate layers were dried over sodium sulfate and concentratedunder reduced pressure to afford crude material which was purified bycolumn chromatography over silica gel obtainingN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-((dimethylamino)methyl)phenyl)-6-(piperidin-1-yl)isonicotinamideas light green solid (0.173 g, 79%).

Analytical Data: LCMS: 474.30 (M+1)⁺; HPLC: 99.15% (@ 254 nm) (R_(t);5.257; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.50(s, 1H), 8.61 (t, 1H, J=4.4 Hz), 8.03 (d, 2H, J=7.6 Hz), 7.52 (s, 1H),7.40 (d, 2H, J=8.4 Hz), 7.13 (s, 1H), 5.87 (s, 1H), 4.32 (d, 2H, J=4Hz), 3.63 (bs, 6H), 2.26 (bs, 6H), 2.18 (s, 3H), 2.11 (s, 3H), 1.59 (bs,6H).

Example 7: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-((dimethylamino)methyl)phenyl)-6-(isopropylamino)isonicotinamide

Step 1: Synthesis of methyl 2-chloro-6-(isopropylamino)isonicotinate

A solution of methyl 2,6-dichloroisonicotinate (1 g, 4.85 mmol),isopropyl amine (0.286 g, 4.85 mmol), Cs₂CO₃ (2.06 g, 6.3 mmol) intoluene (30 mL) was purged with argon for 10 min. Then, Pd(OAc)₂ (0.108g, 0.485 mmol) and BINAP (0.3 g, 0.485 mmol) were added and argon waspurged again for 15 min. Reaction mass was stirred at 80° C. for 6 h. Oncompletion, reaction mass was filtered and residue washed thoroughlywith ethyl acetate. Combined filtrates were concentrated and purified bycolumn over silica gel to obtain pure methyl2-chloro-6-(isopropylamino)isonicotinate (0.3 g, 27.27%).

Step 2: Synthesis of2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(isopropylamino)isonicotinamide

To a solution of methyl 2-chloro-6-(isopropylamino)isonicotinate (0.393g, 1.7 mmol) in ethanol (4 mL), NaOH (0.082 g, 2.06 mmol), water (0.8mL) were added and reaction mass heated at 60° C. for 1 h. Oncompletion, solvent was evaporated under reduced pressure. Residue waswashed with ether and acidified with 1N HCl till pH 8 and then withcitric acid till pH 5-6. Solid obtained was filtered, washed with waterand finally dried under reduced pressure to afford the acid (0.36 g,97%) which was used in next step without further purification. To asolution of this acid (0.36 g, 1.68 mmol) in DMSO (1.5 mL), PyBOP (1.3g, 2.5 mmol) was added and reaction stirred at rt for 15 min. Then3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.383 g, 2.5 mmol) wasadded and reaction stirred overnight. On completion, water was added andaqueous layer extracted with 10% MeOH/DCM. Combined organic layers werewashed with water, dried over sodium sulfate and concentrated to obtaincrude2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(isopropylamino)isonicotinamide(0.58 g, 100%) which was used in next step without further purification.

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-formylphenyl)-6-(isopropylamino)isonicotinamide

To a stirred solution of2-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(isopropylamino)isonicotinamide(0.58 g, 1.67 mmol), boronic acid (0.277 g, 1.84 mmol) in dioxane/watermixture (7 mL+3 mL), Na₂CO₃ (0.64 g, 6.037 mmol) was added and reactionmass purged for 15 min with argon. Then Pd(PPh₃)₄ (0.194 g, 0.168 mmol)was added and argon was purged again for 10 min. Reaction mass washeated at 100° C. for 3 h. On completion, reaction mass filtered throughcelite and celite bed washed with ethyl acetate. Combined filtrates weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column over silica gel to obtainN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-formylphenyl)-6-(isopropylamino)isonicotinamide(0.6 g, 85.7%).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-((dimethylamino)methyl)phenyl)-6-(isopropylamino)isonicotinamide

To a solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-formylphenyl)-6-(isopropylamino)isonicotinamide(0.6 g, 1.44 mmol) in methanol (6 mL), dimethyl amine (7.1 mL, 14.33mmol, 2M solution in THF) and acetic acid (0.086 g, 1.44 mmol) was addedand reaction mass stirred at rt for 1 h. Then sodium cyanoborohydride(0.18 g, 2.8 mmol) was added reaction stirred at rt for 2 h. Oncompletion, solvent was removed under reduced pressure, residue treatedwith water and extracted with ethyl acetate. Combined ethyl acetatelayers were dried over sodium sulfate and concentrated under reducedpressure to afford crude material which was purified by prep HPLCobtaining target molecule Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(4-((dimethylamino)methyl)phenyl)-6-(isopropylamino)isonicotinamideas light yellow solid.

Analytical Data: LCMS: 448.25 (M+1)₊; HPLC: 96.22% (@ 254 nm) (R_(t);4.170, Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (CD₃OD, 400 MHz) δ 8.01 (d,2H, J=8 Hz), 7.67 (d, 2H, J=8 Hz), 7.39 (s, 1H), 7.19 (s, 1H), 6.14 (s,1H), 4.50 (s, 2H), 4.40 (s, 2H), 4.17-4.11 (m, 1H), 2.89 (s, 6H), 2.38(s, 3H), 2.25 (s, 3H), 1.33 (d, 6H, J=6H).

Example 8: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

Step 1: Synthesis of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (2.5 g,10.2 mmol) and dihydro-2H-pyran-4(3H)-one (1.3 g, 13.3 mmol) in methanol(20 mL), acetic acid (0.61 g, 10.2 mmol) was added and the solutionstirred at room temperature for 18 h. Then sodium cyanoborohydride (1.2g, 20.48 mmol) was added at 0° C. and stirring was continued overnightat room temperature. Then, solvent was removed under reduced pressureand crude material was purified by column chromatography to affordmethyl 5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate(2.2 g, 66%).

Step 2: Synthesis of methyl 5-bromo-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate (1.0 g,3.15 mmol) in acetonitrile (15 mL), cesium carbonate (1.97 g, 6.10 mmol)and methyl iodide (2.15 g, 15.27 mmol) were added; resulting solutionwas heated at 80° C. for 20 h. The solution was cooled to roomtemperature, filtered, and the residue was washed with ethyl acetate.The filtrate was concentrated and the product purified by columnchromatography to afford methyl 5-bromo-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzoate (0.82 g, 80%).

Step 3: Synthesis of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzamide

Aqueous NaOH (0.19 g, 4.89 mmol) was added to a solution of methyl5-bromo-2-methyl-3-(methyl (tetrahydro-2H-pyran-4-yl) amino) benzoate(0.82 g, 2.4 mmol) in MeOH (20 mL) and stirred at 60° C. for 1 h.Ethanol was removed under reduced pressure and the solution acidifiedusing dilute HCl to pH 6 and citric acid to pH 4. The product wasextracted with ethyl acetate and the combined organic layers were driedand concentrated to give respective acid (0.70 g). The acid was thendissolved in DMSO (3 mL) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.74 g, 4.89 mmol) was added toit. The reaction mixture was stirred at room temperature for 15 min thenPYBOP (1.9 g, 3.6 mmol) was added to it and stirring was continued forovernight. The solution was poured into ice to obtain a solid, this wasfiltered and washed with acetonitrile followed by purification withcolumn chromatography to afford 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzamide (0.6 g, 54%).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzamide (1 equiv.) and4-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The solutionwas heated at 100° C. for 4 h. The reaction mixture was diluted withwater and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford desiredN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide(0.045 g, 36.9%).

LCMS: 563.00 (M+1)₊; HPLC % 99.26 (@ 254 nm) (R_(t); 3.774; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.17(s, 1H), 8.06 (t, 1H, J=4.8 Hz), 7.82 (s, 1H), 7.29 (s, 1H), 7.11 (s,1H), 5.87 (s, 1H), 4.27 (d, 2H, J=4.8 Hz), 4.21 (t, 2H, J=6.4 Hz), 3.85(d, 2H, J=11.2 Hz), 3.54 (t, 4H), 3.23-3.26 (m, 2H), 2.99 (m, 1H), 2.72(t, 2H, J=6.4 Hz), 2.60 (s, 3H), 2.40 (bs, 4H), 2.20 (s, 3H), 2.16 (s,3H), 2.10 (s, 3H), 1.58-1.59 (m, 4H).

Example 9: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-methyl-1H-pyrazol-4-yl)benzamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzamide (1 equiv.) and1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The solutionwas heated at 100° C. for 4 h. The reaction mixture was diluted withwater and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford desiredN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-methyl-1H-pyrazol-4-yl)benzamide(0.02 g, 20%).

LCMS: 464.30 (M+1)⁺; HPLC % 97.80 (@ 254 nm) (R_(t); 4.286; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.12(s, 1H), 8.06 (t, 1H), 7.81 (s, 1H), 7.28 (s, 1H), 7.10 (s, 1H), 5.85(s, 1H), 4.27 (d, 2H, J=4.8 Hz), 3.83-3.86 (m, 5H), 3.23-3.29 (m, 2H),2.99 (m, 1H), 2.59 (s, 3H), 2.20 (s, 3H), 2.16 (s, 3H), 2.10 (s, 3H),1.58 (m, 4H).

Example 10: Synthesis of3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

Step 1: Synthesis of methyl 5-bromo-3-(cyclohexylamino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5.0 g,20.6 mmol) and cyclohexanone (4.03 g, 41.2 mmol) in methanol (50 mL),acetic acid (0.247 g, 20.6 mmol) was added and reaction stirred at roomtemperature for 3 h. Then sodium cyanoborohydride (1.55 g, 24.6 mmol)was added and reaction stirred overnight. On completion, solvent wasremoved under reduced pressure and crude material was purified by columnchromatography to afford methyl5-bromo-3-(cyclohexylamino)-2-methylbenzoate (2.75 g, 41%).

Step 2: Synthesis of methyl 5-bromo-3-(cyclohexyl (methyl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-3-(cyclohexylamino)-2-methylbenzoate (2.75 g, 8.45 mmol) inacetonitrile (25 mL), cesium carbonate (5.45 g, 16.9 mmol) and methyliodide (6 g, 42.3 mmol) were added; resulting solution was heated at 80°C. for 20 h. On completion, the solution was cooled to room temperatureand filtered, and the residue was washed with ethyl acetate. Thefiltrate was concentrated and then purified by column chromatography toafford methyl 5-bromo-3-(cyclohexyl (methyl) amino)-2-methylbenzoate(2.5 g, 87%).

Step 3: Synthesis of 5-bromo-3-(cyclohexyl (methyl) amino)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-2-methylbenzamide

Aqueous NaOH (0.55 g, 14.7 mmol) was added to a solution of methyl5-bromo-3-(cyclohexyl (methyl) amino)-2-methylbenzoate (2.5 g, 7.35mmol) in MeOH (15 mL) and stirred at 60° C. for 1 h. After completion ofthe reaction, ethanol was removed under reduced pressure and acidifiedusing dilute HCl to pH 6 and citric acid to pH 4. The product wasextracted with ethyl acetate. Combined organic layers were dried andconcentrated to give the respective acid (2.5 g, 87%). The acid was thendissolved in DMSO (20 mL) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2.34 g, 15.1 mmol) was added toit. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (5.85 g, 11.05 mmol) was added to it and stirring wascontinued for overnight. Then the reaction was poured into ice to obtaina solid which was collected by filtration and washed with acetonitrile.Column purification on silica provided 5-bromo-3-(cyclohexyl (methyl)amino)-N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (1.5 g, 44.19%).

Step 4: Synthesis of3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

To a stirred solution of 5-bromo-3-(cyclohexyl (methyl) amino)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-2-methylbenzamide (1equiv.) and1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd (PPh₃)₄(0.1 equiv.) was added and argon was purged again for 10 min. Thereaction was heated at 100° C. for 4 h. After cooling, the reactionmixture was diluted with water and extracted with 10% MeOH/DCM. Thecombined organic layers were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford crude product. Purification by columnchromatography over silica gel afforded the title compound (0.02 g,20%).

LCMS: 462.40 (M+1)₊; HPLC % 88.48 (@ 254 nm) (R_(t); 4.683; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.11(s, 1H), 8.06 (t, 1H), 7.79 (s, 1H), 7.22 (s, 1H), 7.06 (s, 1H), 5.85(s, 1H), 4.26 (d, 2H, J=4 Hz), 3.83 (s, 3H), 2.71 (t, 1H), 2.60 (s, 3H),2.20 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H), 1.69 (m, 4H), 1.53-1.55 (m,1H), 1.39-1.41 (m, 2H), 1.06-1.19 (m, 3H).

Example 11: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-5-(methyl(piperidin-4-yl)amino)-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of tert-butyl 4-((5-bromo-3-(methoxycarbonyl) amino)piperidine-1-carboxylate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5.0 g,20.6 mmol) and tert-butyl 4-oxopiperidine-1-carboxylate (8.2 g, 41.1mmol) in methanol (50 mL), acetic acid (1.2 g, 20.6 mmol) was added andthe reaction stirred at room temperature for 8 h. Then sodiumcyanoborohydride (1.55 g, 24.6 mmol) was added at 0° C. and the reactionstirred overnight at room temperature. The solvent was removed underreduced pressure and the product was purified by column chromatographyon silica gel to afford tert-butyl 4-((5-bromo-3-(methoxycarbonyl)amino) piperidine-1-carboxylate (5.0 g, 57%).

Step 2: Synthesis of tert-butyl4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl (methyl) amino)piperidine-1-carboxylate

To a stirred solution of tert-butyl 4-((5-bromo-3-(methoxycarbonyl)amino) piperidine-1-carboxylate (3.0 g, 7.06 mmol) in acetonitrile (25mL), cesium carbonate (4.57 g, 14.1 mmol) and methyl iodide (5.0 g, 35.2mmol) were added. The reaction was heated to 80° C. for 20 h. Then thereaction was cooled to room temperature and filtered, washing with ethylacetate. The filtrate was concentrated and the product purified bycolumn chromatography on silica gel to afford tert-butyl4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl (methyl) amino)piperidine-1-carboxylate (2.5 g, 81%).

Step 3: Synthesis of tert-butyl 4-((5-bromo-3-(((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl) carbamoyl)-2-methylphenyl) (methyl)amino) piperidin-1-carboxylate

Aqueous NaOH (0.37 g, 9.38 mmol) was added to a solution of tert-butyl4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl (methyl) amino)piperidine-1-carboxylate (2.0 g, 4.69 mmol) in MeOH (20 mL) and stirredat 60° C. for 1 h. After completion of the reaction, ethanol was removedunder reduced pressure and the solution acidified using dilute HCl to pH6 and citric acid to pH 4. The product was extracted using ethylacetate. The combined organic layers were dried and concentrated to givethe respective acid (1.7 g, 90%). The acid was then dissolved in DMSO(10 mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (1.42 g, 9.38mmol) was added to it. The reaction mixture was stirred at roomtemperature for 15 min before PYBOP (3.66 g, 7.04 mmol) was added to itand stirring was continued for overnight. After completion, reactionmass was poured into ice to obtain solid, this was filtered and washedwith acetonitrile followed by purification with column chromatography toafford tert-butyl 4-((5-bromo-3-(((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl) carbamoyl)-2-methylphenyl) (methyl)amino) piperidin-1-carboxylate (1.3 g, 50%).

Step 4: Synthesis of tert-butyl4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)piperidine-1-carboxylate

To a stirred solution of tert-butyl 4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)carbamoyl)-2-methylphenyl) (methyl) amino) piperidin-1-carboxylate (1equiv.) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine (1.2equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) wasadded and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The reactionwas heated at 100° C. for 5 h. After cooling, the reaction mixture wasdiluted with water, and the product was extracted with 10% MeOH/DCM. Thecombined organic layers were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)piperidine-1-carboxylate

Step 5: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-5-(methyl(piperidin-4-yl)amino)-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A stirred solution of tert-butyl4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)piperidine-1-carboxylate(1 mmol) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added toit. The reaction was stirred at room temperature for 1 h. On completion,the solution was concentrated to dryness. The residue was purified bysolvent washings to afford the title compound (0.07 g, 86%).

LCMS: 558.45 (M+1)₊; HPLC % 98.81 (@ 254 nm) (R_(t); 4.009; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 10.1(bs, 1H), 8.51 (d, 1H), 8.16 (t, 2H), 7.77 (d, 2H, J=8 Hz), 7.57 (d, 2H,J=8 Hz), 7.42 (s, 1H), 7.26 (s, 1H), 5.86 (s, 1H), 4.33 (bs, 2H), 4.29(d, 2H, J=19.2 HZ), 3.96 (m, 2H), 3.25 (m, 4H), 3.15 (m, 4H), 2.89-2.91(m, 2H), 2.64 (s, 3H), 2.26 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.81(m, 4H).

Example 12: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-5-(methyl(tetrahydro-2H-pyran-4-yl)amino)-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzamide (1 equiv.) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine (1.2equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) wasadded and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The solutionwas heated at 100° C. for 4 h. The reaction mixture was diluted withwater and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford the title compound (0.065 g, 55%). LCMS: 559.35 (M+1)⁺;HPLC % 99.26 (@ 254 nm) (R_(t); 3.983; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 L, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.15 (t, 1H), 7.58 (d,2H, J=8 Hz), 7.36 (d, 3H, J=8.4 Hz), 7.18 (s, 1H), 5.85 (s, 1H), 4.28(d, 2H, J=4.8 Hz), 3.84 (d, 2H, J=11.2 Hz), 3.57 (m, 3H), 3.48 (m, 3H),3.24 (m, 2H), 3.40 (m, 1H), 2.63 (s, 3H), 2.36 (m, 4H), 2.23 (s, 3H),2.20 (s, 3H), 2.10 (s, 3H), 1.60 (m, 4H).

Example 13: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-5-(methyl(tetrahydro-2H-pyran-4-yl)amino)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzamide (1 equiv.) andN,N-dimethyl-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanamine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The solutionwas heated at 100° C. for 4 h. The reaction mixture was diluted withwater and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford the title compound (0.01 g, 9%). LCMS: 517.30 (M+1)+; HPLC% 98.12 (@ 254 nm) (R_(t); 3.972; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient:5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.16 (t, 1H), 7.58 (d, 2H, J=8 Hz),7.34-7.36 (m, 2H), 7.18 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4 Hz),3.84 (d, 2H, J=10.8 Hz), 3.42 (s, 2H), 3.02 (m, 2H), 2.66 (m, 1H), 2.63(s, 3H), 2.50 (3H merged in solvent peak), 2.23 (s, 3H), 2.20 (s, 3H),2.16 (s, 3H), 2.10 (s, 3H), 1.60 (m, 4H).

Example 14: Synthesis of5-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.) and (4-(morpholinomethyl) phenyl)boronic acid (1.2 equiv.) indioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) was added and thesolution was purged with argon for 15 min. Then Pd (PPh₃)₄ (0.1 equiv.)was added and solution was purged again for 10 min. Reaction mixture washeated at 100° C. for 4 h. On completion, the mixture was diluted withwater and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford the title compound (0.070 g, 29% yield). LCMS: 557.40(M+1)⁺; HPLC % 98.83 (@ 254 nm) (R_(t); 4.303; Method: Column: YMC ODS-A150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.15 (t, 1H, J=4 Hz),7.56 (d, 2H, J=7.6 Hz), 7.36 (d, 2H, J=8 Hz), 7.28 (s, 1H), 7.13 (s,1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.57 (m, 4H), 3.48 (s, 2H),2.74 (t, 1H), 2.64 (s, 3H), 2.36 (m, 4H), 2.20 (s, 6H), 2.10 (s, 3H),1.69-1.71 (m, 3H), 1.53-1.56 (m, 2H), 1.41-1.44 (m, 2H), 1.10-1.23 (m,3H).

Example 15: Synthesis of3-(Cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

To a stirred solution of5-bromo-3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.) and (1-(2-morpholinoethyl)-1H-pyrazol-4-yl)boronic acid (1.2equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) wasadded and the solution was purged with argon for 15 min. Then Pd (PPh₃)₄(0.1 equiv.) was added and solution was purged again for 10 min.Reaction mixture was heated at 100° C. for 4 h. On completion, themixture was diluted with water and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over Na₂SO₄ and solvent removed under reducedpressure to afford crude material which was purified by columnchromatography over silica gel to afford the title compound (0.06 g, 25%yield). LCMS: 561.35 (M+1)⁺; HPLC % 96.87 (@ 254 nm) (R_(t); 4.209;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.44 (s, 1H),8.15 (s, 1H), 8.06 (t, 1H), 7.81 (s, 1H), 7.22 (s, 1H), 7.06 (s, 1H),5.85 (s, 1H), 4.28 (d, 2H, J=4.8 Hz), 4.21 (t, 2H, J=6 Hz), 3.54 (m,4H), 2.72 (t, 2H, J=6.8 Hz), 2.61 (s, 3H), 2.40 (m, 4H), 2.20 (s, 3H),2.14 (s, 3H), 2.10 (s, 3H), 1.70 (m, 4H), 1.53-1.56 (m, 3H), 1.10-1.23(m, 4H).

Example 16: Synthesis of5-(Cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.) and (4-((dimethylamino) methyl)phenyl)boronic acid (1.2equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) wasadded and the solution was purged with argon for 15 min. Then Pd (PPh₃)₄(0.1 equiv.) was added and solution was purged again for 10 min.Reaction mixture was heated at 100° C. for 4 h. On completion, themixture was diluted with water and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over Na₂SO₄ and solvent removed under reducedpressure to afford crude material which was purified by columnchromatography over silica gel to afford the title compound (0.065 g,29% yield). LCMS: 515.40 (M+1)⁺; HPLC % 96.73 (@ 254 nm) (R_(t); 4.362;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H),8.16 (t, 1H), 7.64 (d, 2H, J=6.8 Hz), 7.45 (d, 2H), 7.30 (s, 1H), 7.16(s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 2.75 (t, 1H), 2.65 (s,3H), 2.32-2.42 (m, 6H), 2.20 (s, 6H), 2.10 (s, 3H), 1.69 (m, 4H),1.53-1.56 (m, 1H), 1.42-1.45 (m, 2H), 1.10-1.23 (m, 4H). [1H merged insolvent peak].

Example 17: Synthesis of3-(Cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Synthesis of3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide

To a stirred solution of5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.5 g, 1.12 mmol) and (6-formylpyridin-3-yl)boronic acid (0.39 g, 1.68mmol) in dioxane/water mixture (15 mL+3 mL), Na₂CO₃ (0.42 g, 4.09 mmol)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄(0.130 g, 0.112 mmol) was added the mixture was purged again for 10 min.Reaction mass was heated at 100° C. for 4 h. On completion, reactionmixture was diluted with water and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over Na₂SO₄ and solvent removed under reducedpressure to afford crude material which was purified by columnchromatography over silica gel to afford the title compound (0.35 g, 66%yield).

Step 2: Synthesis of3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

To a stirred solution of compound3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(1 equiv.) and morpholine (5 equiv.) in methanol (10 mL), acetic acid (2equiv.) was added and reaction stirred at room temperature for 18 h.Then sodium cyanoborohydride (2.5 equiv.) was added at 0° C. andreaction stirred overnight at room temperature. On completion, solventwas removed under reduced pressure and crude material was purified bycolumn chromatography to afford compound and crude material which waspurified by preparative HPLC giving the title compound as a TFA salt,(0.022 g, 22%). LCMS: 544.35 (M+1)⁺; HPLC % 98.42 (@ 254 nm) (R_(t);4.143; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.75 (s, 1H), 8.17 (t, 1H), 8.01 (d, 1H, J=7.6) 7.50 (d, 1H,J=7.6 Hz), 7.42 (s, 1H), 7.22 (s, 1H), 7.06 (s, 1H), 5.85 (s, 1H), 4.28(d, 2H), 3.59-3.61 (m, 8H), 3.35-3.37 (m, 2H), 2.66 (s, 1H), 2.55 (s,3H), 2.24 (s, 3H), 2.19 (s, 3H), 2.10 (s, 3H), 1.72 (m, 2H), 1.61 (m,2H), 1.48 (m, 4H).

Example 18: Synthesis of3-(Cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

A solution of5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.), (1-(2-morpholinoethyl)-1H-pyrazol-4-yl)boronic acid (1.2equiv.) and Pd (PPh₃)₄ (0.1 equiv.) in 1, 4-dioxane (4 mL) was purgedwith argon for 10 min. Then, 2 M Na₂CO₃ solution (3.6 equiv.) was addedto it and the mixture was purged again for 10 min. The reaction mixturewas stirred at 100° C. for 2 h. After reaction completion, water wasadded to it and extraction was carried out using DCM. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography over silica gel(60-120 mesh size) to afford 3 the title compound (0.08 g, 66%). LCMS:547.35 (M+1)⁺; HPLC % 97.60 (@ 254 nm) (R_(t); 4.071; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.44 (s, 1H), 8.17 (s, 1H), 8.05 (t,1H), 7.81 (s, 1H), 7.30 (s, 1H), 7.10 (s, 1H), 5.85 (s, 1H), 4.26 (d,2H, J=4 Hz), 4.20 (d, 2H, J=6.4 Hz), 3.49-3.53 (m, 6H), 2.72 (t, 2H),2.40 (bs, 6H), 2.20 (s, 3H), 2.17 (s, 3H), 2.10 (s, 3H), 1.61-1.70 (m,4H), 1.42-1.50 (m, 4H).

Example 19: Synthesis of3-(Cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

A solution of5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(1 equiv.), (1-methyl-1H-pyrazol-4-yl)boronic acid (1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1, 4-dioxane (4 mL) was purged with argon for 10min. Then, 2 M Na₂CO₃ solution (3.6 equiv.) was added to it and themixture was purged again for 10 min. The reaction mixture was stirred at100° C. for 2 h. After reaction completion, water was added andextraction was carried out using DCM. The combined organic layers werewashed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford the title compound (0.07 g, 70%) LCMS: 448.25 (M+1)⁺; HPLC %98.34 (@ 254 nm) (R_(t); 4.578; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient:5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.44 (s, 1H), 8.11 (s, 1H), 8.05 (t, 1H), 7.80 (s,1H), 7.29 (s, 1H), 7.09 (s, 1H), 5.85 (s, 1H), 4.26 (d, 2H, J=3.2 Hz),3.83 (s, 3H), 3.49 (m, 1H), 2.20 (s, 3H), 2.16 (s, 3H), 2.10 (s, 3H),1.69 (m, 2H), 1.60 (m, 2H), 1.42-1.49 (m, 4H). [3H merged in solventpeak].

Example 20: Synthesis of5-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic acid

To a stirred solution of 2-methyl-3-nitrobenzoic acid (50 g, 276.2 mmol)in conc. H₂SO₄ (200 mL) was added1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8 mmol)portion wise at room temperature and the reaction mixture was stirred atroom temperature for 5 h. On completion, the reaction mixture was pouredonto ice cold water, the resulting precipitate was filtered, the residuewas washed with water and dried under vacuum to give5-bromo-2-methyl-3-nitrobenzoic acid (71.7 g, 99.9%) which was useddirectly in the next step.

Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate

To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid (287 g,1103 mmol) in DMF (150 mL) was added sodium carbonate (468 g, 4415 mmol)and methyl iodide (626.63 g, 4415 mmol). The resulting reaction mixturewas heated at 60° C. for 8 h. On completion, the precipitated solid wascollected by filtration, the residue washed with diethyl ether (5times). The combined organic layers were dried, concentrated underreduced pressure to give methyl 5-bromo-2-methyl-3-nitrobenzoate (302 g,99%) which was used directly in the next step.

Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate

To a stirred solution of methyl 5-bromo-2-methyl-3-nitrobenzoate (150 g,544 mmol) in ethanol (750 mL) was added ammonium chloride (150 g, 2777mmol) dissolved in water (750 mL) and iron powder (93.3 g, 1636 mmol)with stirring. The resulting reaction mixture was heated at 80° C. for 7h. On completion, the reaction mixture was filtered through celite; theresidue was washed with water and ethyl acetate, filtrate was extractedwith ethyl acetate. The combined organic layers were dried, concentratedunder reduced pressure to give methyl 3-amino-5-bromo-2-methylbenzoatewhich was used directly in the next step.

Step 4: Synthesis of methyl5-bromo-3-((4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-amino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5 g,20.57 mmol) and tert-butyl (4-oxocyclohexyl)carbamate (5.6 g, 26.7 mmol)in methanol (50 mL) was added acetic acid (1.2 g, 20.57 mmol) and thereaction mixture stirred at room temperature for 8 h. Then sodiumcyanoborohydride (1.6 g, 26.74 mmol) was added at 0° C. and the reactionstirred overnight. On completion, solvent was removed under reducedpressure and the crude material was purified by column chromatographytwice eluting with ethyl acetate: hexane to afford methyl5-bromo-3-((4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-amino)-2-methylbenzoate4 g (44%) of non-polar isomer (cis isomer, contaminated with starting)and 3 g (33%) of pure polar isomer (trans isomer).

Step 5: Synthesis of methyl5-bromo-3-((1s,4s)-(4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(methyl)-amino)-2-methylbenzoate

To a stirred solution of the cis isomer of methyl5-bromo-3-((4-((tert-butoxycarbonyl) amino) cyclohexyl)amino)-2-methylbenzoate (4 g, 9.09 mmol) in acetonitrile (50 mL) wasadded cesium carbonate (5.9 g, 18.18 mmol) and methyl iodide (6.45 g,45.45 mmol). The resulting reaction mixture was heated at 80° C. for 7h. On completion, the reaction mixture was cooled to room temperatureand filtered, the residue was washed with ethyl acetate and the filtrateconcentrated then purified by column chromatography to give 4.0 g (44%)of the less-polar cis-isomer, methyl5-bromo-3-(((1s,4s)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-amino)-2-methylbenzoate)and 3.0 g (33%) of more polar trans-isomer,methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-amino)-2-methylbenzoate.

Step 6: Synthesis of tert-butyl(1s,4s)-(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-2-methylphenyl)-(methyl)-amino)-cyclohexyl)carbamate

Aqueous NaOH (0.23 g, 5.72 mmol) was added to a solution of methyl5-bromo-3-(((1s,4s)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(methyl)-amino)-2-methylbenzoate(1.3 g, 2.86 mmol) in MeOH (20 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand acidified using dilute HCl up to pH 6 and adjusted to pH 4 withcitric acid. Extraction was carried out using ethyl acetate. Thecombined organic layers were dried concentrated to give the crude acid(1.13 g, 90.1%).

The acid (1.13 g, 2.57 mmol) was then dissolved in DMSO (10 mL) and3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.87 g, 5.72 mmol) wasadded. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (2.23 g, 4.28 mmol) was added and stirring was continuedovernight. After completion of the reaction, the reaction mixture waspoured into ice to obtain a solid, this was filtered and washed withacetonitrile followed by purification with column chromatography toafford tert-butyl(1s,4s)-(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-2-methylphenyl)-(methyl)-amino)-cyclohexyl)carbamate(0.8 g, 48.7%).

Step 7: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide

To a stirred solution of tert-butyl(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-2-methylphenyl)-(methyl)-amino)-cyclohexyl)-carbamate(0.8 g, 1.39 mmol) in DCM (25 mL) at 0° C. was added TFA (5 mL). Thereaction mixture was stirred at room temperature for 1 h. On completion,the reaction mixture was concentrated to dryness. The residue wasbasified with aqueous sodium bicarbonate to pH 8 and the aqueous layerextracted with 20% MeOH/DCM. The combined organic layers were dried oversodium sulfate and concentrated to afford3-(((1s,4s)-4-aminocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide(600 mg, 90.9%).

Step 8: Synthesis of3-((1s,4s)-(4-acetamidocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide

To a stirred solution of3-((4-aminocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide(0.275, 0.580 mmol) in DMF (5 mL), was added EDCI.HCl (0.168 g, 0.870mmol), HOBt (0.078 g, 0.58 mmol) and acetic acid (0.07 g, 1.16 mmol),the reaction mixture was stirred at room temperature for 18 h. Oncompletion, water was added and the organics extracted with 10%MeOH/DCM. The combined organic layers were dried, concentrated givingcrude material which then purified by column chromatography to afford3-(((1s,4s)-4-acetamidocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide(0.25 g, 83.6%).

Step 9: Synthesis of5-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of3-((4-acetamidocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide(1 equiv.) and 4-(4-(4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)benzyl) morpholine (1.2 equiv.) in dioxane/water mixture (5 mL+1 mL) wasadded Na₂CO₃ (3.6 equiv.) and the solution purged with argon for 15 min.Then Pd(PPh₃)₄ (0.1 equiv.) was added and argon was purged again for 10min. The reaction mixture was heated at 100° C. for 4 h. On completion,the reaction mixture was diluted with water and extracted with 10%MeOH/DCM. The combined organic layers were dried over Na₂SO₄ and thesolvent removed under reduced pressure to afford crude material whichwas purified by column chromatography over silica gel to afford thetitle compound (0.06 g, 50.8%). LCMS: 614.40 (M+1)⁺; HPLC % 99.44 (@ 254nm) (R_(t); 3.948; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.45 (s, 1H), 8.17 (t, 1H), 7.76 (d, 1H, J=7.2 Hz), 7.55 (d, 2H,J=7.6 Hz), 7.36 (d, 3H, J=8 Hz), 7.16 (s, 1H), 5.85 (s, 1H), 4.28 (d,2H, J=4.4 Hz), 3.71 (bs, 1H), 3.57 (m, 4H), 3.47 (s, 2H), 2.98 (m, 1H),2.59 (s, 3H), 2.36 (m, 4H), 2.26 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H),1.74-1.81 (m, 5H), 1.49-1.56 (m, 3H), 1.40-1.48 (m, 3H).

Example 21:5-(((1r,4r)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamideprepared in analogous fashion as example 20 from trans-isomer, methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-amino)-2-methylbenzoateintermediate described in Example 20

Analytical Data of 1258-Trans: LCMS: 614.40 (M+1)⁺; HPLC % 99.64 (@ 254nm) (R_(t); 3.917; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.45 (s, 1H), 8.16 (t, 1H), 7.76 (d, 1H, J=7.6 Hz), 7.57 (d, 2H,J=7.2 Hz), 7.36 (d, 2H, J=7.6 Hz), 7.29 (s, 1H), 7.14 (s, 1H), 5.85 (s,1H), 4.28 (d, 2H), 3.57 (bs, 5H), 3.48 (m, 2H), 2.71 (m, 1H), 2.64 (s,3H), 2.36 (m, 4H), 2.20 (s, 6H), 2.10 (s, 3H), 1.68-1.81 (m, 7H),1.51-1.53 (m, 2H), 1.10-1.13 (m, 2H).

Example 22: Synthesis of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

To a stirred solution of3-(((1s,4s)-4-acetamidocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide(1 equiv.) and1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL) was added Na₂CO₃ (3.6equiv.) and the solution purged with argon for 15 min. Then Pd(PPh₃)₄(0.1 equiv.) was added and argon was purged again for 10 min. Thereaction mixture was heated at 100° C. for 4 h. On completion, thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic layers were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford crude material which waspurified by column chromatography over silica gel to afford the titlecompound (0.02 g, 20%). LCMS: 519.40 (M+1)⁺; HPLC % 96.24 (@ 254 nm)(R_(t); 4.247; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.44 (s, 1H), 8.10 (s, 1H), 8.07 (t, 1H), 7.79 (s, 1H), 7.75 (d, 1H,J=7.2 Hz), 7.27 (s, 1H), 7.09 (s, 1H), 5.86 (s, 1H), 4.27 (d, 2H, J=4.8Hz), 3.83 (s, 3H), 3.69 (bs, 1H), 2.96 (m, 1H), 2.56 (s, 3H), 2.20 (s,6H), 2.10 (s, 3H), 1.81 (s, 3H), 1.74-1.76 (m, 2H), 1.54 (m, 2H),1.36-1.46 (m 4H).

Example 23 Synthesis of3-(((1r,4r)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

Prepared in prepared in analogous fashion as example 22 (0.06 g, 40%).LCMS: 519.30 (M+1)⁺; HPLC % 98.21 (@ 254 nm) (R_(t); 4.155; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.12(s, 1H), 8.07 (t, 1H), 7.80 (s, 1H), 7.66 (d, 1H, J=7.2 Hz), 7.23 (s,1H), 7.07 (s, 1H), 5.86 (s, 1H), 4.26 (d, 2H, J=2.8 Hz), 3.83 (s, 3H),3.44 (m, 1H), 2.66-2.69 (m, 1H), 2.61 (s, 3H), 2.20 (s, 3H), 2.13 (s,3H), 2.10 (s, 3H), 1.78-1.80 (m, 2H), 1.74 (s, 3H), 1.67-1.70 (m, 2H),1.48-1.51 (m 2H), 1.10-1.13 (m, 2H).

Example 24: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(piperidin-4-yl)amino)-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Synthesis of tert-butyl 4-((3-(((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl) (methyl) amino)piperidine-1-carboxylate

Tert-butyl 4-((5-bromo-3-(((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl) carbamoyl)-2-methylphenyl) (methyl)amino) piperidin-1-carboxylate (0.5 g, 0.892 mmol),(6-formylpyridin-3-yl)boronic acid (0.31 g, 1.33 mmol) and Pd(PPh₃)₄(0.103 g, 0.082 mmol) in 1,4-dioxane (10 mL) was purged with argon for10 min. Then, 2 M Na₂CO₃ solution (0.34 g, 3.21 mmol) was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 2 h. After completion of the reaction, waterwas added to it and extraction was carried out using 5% MeOH in DCM. Thecombined organic layers were washed with water, dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography over silica gel(60-120 mesh size) to afford tert-butyl4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)piperidine-1-carboxylate(0.40 g, 87.9%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(piperidin-4-yl)amino)-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

To a stirred solution of tert-butyl4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)piperidine-1-carboxylate(1 equiv.) and morpholine (5 equiv.) in methanol (5 mL for 0.3 mmol),acetic acid (1 equiv.) was added and reaction stirred at roomtemperature for 4 h. Then reducing agent NaBH₃CN (1 equiv.) was addedand reaction stirred overnight. On completion, solvent was removed underreduced pressure and residue purified by column chromatography oversilica gel affording desired tert-butyl4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)phenyl)(methyl)amino)piperidine-1-carboxylate.This compound was then dissolved in DCM (5 mL) and cooled to 0° C. TFA(2 mL) was added to it. Reaction mixture was stirred at room temperaturefor 1 h. On completion, reaction was concentrated to dryness. Residuewas purified by solvent washings to afford the title compound (0.1 g,65.78%). LCMS: 559.35 (M+1)⁺; HPLC: 95.60% (@ 254 nm) (R_(t); 3.906;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H),8.96 (s, 1H), 8.67 (m, 1H), 8.22 (d, 2H, J=8 Hz), 8.17 (t, 1H), 7.61 (d,1H, J=8 Hz), 7.48 (s, 1H), 7.32 (s, 1H), 5.87 (s, 1H), 4.52 (s, 2H),4.29 (d, 2H, J=4.4 Hz), 3.84 (bs, 4H), 3.26 (bs, 6H), 3.16 (t, 1H),2.89-2.91 (m, 2H), 2.64 (s, 3H), 2.26 (s, 3H), 2.21 (s, 3H), 2.10 (s,3H), 1.81 (bs, 4H).

Example 25: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(piperidin-4-yl)amino)-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

To a stirred solution of tert-butyl 4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)carbamoyl)-2-methylphenyl) (methyl) amino) piperidin-1-carboxylate (1equiv.) and4-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The reactionwas heated at 100° C. for 5 h. After cooling, the reaction mixture wasdiluted with water, and the product was extracted with 10% MeOH/DCM. Thecombined organic layers were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)phenyl)(methyl)amino)piperidine-1-carboxylate.A stirred solution of this compound (1 mmol) in DCM (5 mL) was cooled to0° C. and TFA (2 mL) was added to it. The reaction was stirred at roomtemperature for 1 h. On completion, the solution was concentrated todryness. The residue was purified by solvent washings to afford thetitle compound (0.06 g, 89%). LCMS: 562.40 (M+1)⁺; HPLC: 99.01% (@ 254nm) (R_(t); 3.838; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.46 (s, 1H), 8.52 (s, 1H), 8.26 (s, 1H), 8.23 (m, 1H), 8.05 (t, 1H),8.00 (s, 1H), 7.34 (s, 1H), 7.16 (s, 1H), 5.87 (s, 1H), 4.53 (t, 2H),4.27 (d, 2H, J=3.6 Hz), 3.25 (m, 4H), 3.10-3.16 (m, 4H), 2.87 (m, 2H),2.60 (s, 3H), 2.20 (s, 3H), 2.18 (s, 3H), 2.11 (s, 3H), 1.79 (bs, 4H).[5H merged in solvent peak]

Example 26: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(piperidin-4-yl)amino)-5-(1-methyl-1H-pyrazol-4-yl)benzamide

To a stirred solution of tert-butyl 4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)carbamoyl)-2-methylphenyl) (methyl) amino) piperidin-1-carboxylate (1equiv.) and1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The reactionwas heated at 100° C. for 5 h. After cooling, the reaction mixture wasdiluted with water, and the product was extracted with 10% MeOH/DCM. Thecombined organic layers were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)phenyl)(methyl)amino)piperidine-1-carboxylate.A stirred solution of this compound (1 mmol) in DCM (5 mL) was cooled to0° C. and TFA (2 mL) was added to it. The reaction was stirred at roomtemperature for 1 h. On completion, the solution was concentrated todryness. The residue was purified by solvent washings to afford thetitle compound (0.07 g, 87%). LCMS: 463.30 (M+1)⁺; HPLC: 98.02% (@ 254nm) (R_(t); 4.145; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.46 (s, 1H), 8.47 (bs, 1H), 8.12 (s, 2H), 8.05 (s, 1H), 7.83 (s,1H), 7.32 (s, 1H), 7.14 (s, 1H), 5.86 (s, 1H), 4.28 (m, 2H), 3.84 (s,3H), 3.24-3.27 (m, 2H), 3.11 (bs, 1H), 2.87-2.89 (m, 2H), 2.59 (s, 3H),2.20 (s, 3H), 2.18 (s, 3H), 2.10 (s, 3H), 1.77-1.80 (m, 4H).

Example 27: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-5-(methyl(piperidin-4-yl)amino)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of tert-butyl 4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)carbamoyl)-2-methylphenyl) (methyl) amino) piperidin-1-carboxylate (1equiv.) andN,N-dimethyl-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanamine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. The reactionwas heated at 100° C. for 5 h. After cooling, the reaction mixture wasdiluted with water, and the product was extracted with 10% MeOH/DCM. Thecombined organic layers were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(methyl)amino)piperidine-1-carboxylate.A stirred solution of this compound (1 mmol) in DCM (5 mL) was cooled to0° C. and TFA (2 mL) was added to it. The reaction was stirred at roomtemperature for 1 h. On completion, the solution was concentrated todryness. The residue was purified by solvent washings to afford thetitle compound (0.06 g, 90%). LCMS: 516.35 (M+1)⁺; HPLC: 98.28% (@ 254nm) (R_(t); 3.930; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.46 (s, 1H), 9.82 (bs, 1H), 8.51 (bs, 1H), 8.17 (s, 2H), 7.77 (d,2H, J=7.2 Hz), 7.55 (d, 2H, J=7.6 Hz), 7.43 (s, 1H), 7.27 (s, 1H), 5.86(s, 1H), 4.30 (m, 4H), 3.25 (4H merged in solvent peak), 2.88-2.91 (m,1H), 2.75 (s, 6H), 2.64 (s, 3H), 2.25 (s, 3H), 2.20 (s, 3H), 2.10 (s,3H), 1.81 (m, 4H).

Example 28: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl) amino) benzamide (0.4 g, 0.86 mmol) and(6-formylpyridin-3-yl)boronic acid (0.3 g, 1.29 mmol) in dioxane/watermixture (10 mL+2 mL), Na₂CO₃ (0.32 g, 3.09 mmol) was added and solutionpurged with argon for 15 min. Then Pd(PPh₃)₄ (0.092 g, 0.086 mmol) wasadded and argon was purged again for 10 min. The reaction mixture washeated at 100° C. for 6 h. On completion, reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to affordN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide(0.28 g, 66%).

Step 2:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide(1 equiv.) and morpholine (5 equiv.) in methanol (10 mL), acetic acid (2equiv.) was added and reaction stirred at room temperature for 18 h.Then sodium cyanoborohydride (2.5 equiv.) was added at 0° C. andreaction stirred overnight at room temperature. On completion, solventwas removed under reduced pressure and crude material was purified bycolumn chromatography to afford the title compound (0.08 g, 70%). LCMS:560.30 (M+1)⁺; HPLC: 99.22% (@ 254 nm) (R_(t); 3.944; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.76 (s, 1H), 8.17 (t,1H), 8.02 (d, 1H, J=7.6 Hz), 7.50 (d, 1H, J=8 Hz), 7.41 (s, 1H), 7.23(s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.8 Hz), 3.85 (d, 2H, J=11.2 Hz),3.61 (s, 3H), 3.59-3.60 (m, 3H), 3.24-3.29 (m, 2H), 3.02-3.05 (m, 1H),2.64 (s, 3H), 2.42 (bs, 4H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H),1.61 (bs, 4H).

Example 29: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-(hydroxymethyl)pyridin-3-yl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide(1 equiv.) and dimethylamine (5 equiv.) in methanol (10 mL), acetic acid(2 equiv.) was added and reaction stirred at room temperature for 18 h.Then sodium cyanoborohydride (2.5 equiv.) was added at 0° C. andreaction stirred overnight at room temperature. On completion, solventwas removed under reduced pressure and crude material was purified bycolumn chromatography to afford the title compound. LCMS: 491.25 (M+1)⁺;HPLC: 99.58% (@254 nm) (R_(t); 3.984; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47 (s, 1H), 8.75 (s, 1H), 8.19 (t,1H), 8.05 (d, 1H, J=8.4 Hz), 7.52 (d, 1H, J=8.4 Hz), 7.41 (s, 1H), 7.24(s, 1H), 5.86 (s, 1H), 5.44 (t, 1H, J=5.6 Hz), 4.59 (d, 2H, J=5.6 Hz),4.28 (d, 2H, J=4 Hz), 3.85 (d, 2H, J=10.4 Hz), 3.32 (2H merged insolvent peak), 3.03 (m, 1H), 2.64 (s, 3H), 2.24 (s, 3H), 2.20 (s, 3H),2.10 (s, 3H), 1.61 (bs, 4H).

Example 30: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide(1 equiv.) and dimethylamine (5 equiv.) in methanol (10 mL), acetic acid(2 equiv.) was added and reaction stirred at room temperature for 18 h.Then sodium cyanoborohydride (2.5 equiv.) was added at 0° C. andreaction stirred overnight at room temperature. On completion, solventwas removed under reduced pressure and crude material was purified bycolumn chromatography to afford the title compound (0.03 g, 26%). LCMS:518.25 (M+1)⁺; HPLC: 89.16% (@ 254 nm) (R_(t); 3.982; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.81 (s, 1H), 8.18 (t,1H), 8.08 (d, 1H, J=8 Hz), 7.52 (d, 1H, J=8 Hz), 7.43 (s, 1H), 7.26 (s,1H), 5.86 (s, 1H), 4.28 (d, 2H, J=4.8 Hz), 3.83-3.86 (m, 4H), 3.32 (2Hmerged in solvent peak), 3.03 (m, 1H), 2.64 (s, 3H), 2.50 (3H merged insolvent peak), 2.40 (bs, 3H), 2.24 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H),1.60 (bs, 4H).

Example 31: Synthesis of3-(Cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Synthesis of3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide

To a stirred solution of bromo compound5-bromo-3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.6 g, 1.30 mmol) and (6-formylpyridin-3-yl)boronic acid (0.450 g, 1.95mmol) in dioxane/water mixture (8 mL+2 mL), Na₂CO₃ (0.498 g, 4.5 mmol)was added and solution purged with argon for 15 min. Then Pd (PPh₃)₄(0.15 g, 0.129 mmol) was added and the mixture was purged again for 10min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM.Combined organic layers were dried over Na₂SO₄ and solvent removed underreduced pressure to afford crude material which was purified by columnchromatography over silica gel to afford3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(0.525 g, 83%).

Step 2: Synthesis of3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

To a stirred solution of compound3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(1 equiv.) and morpholine (5 equiv.) in methanol (10 mL), acetic acid (2equiv.) was added and reaction stirred at room temperature for 8 h. Thensodium cyanoborohydride (2.5 equiv.) was added at 0° C. and reactionstirred overnight at room temperature. On completion, solvent wasremoved under reduced pressure and crude material was purified by columnchromatography to afford3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide(0.089 g, 53% yield). LCMS: 558.35 (M+1)⁺; HPLC: 96.52% (@ 254 nm)(R_(t); 4.375; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (s, 1H), 8.75 (s, 1H), 8.18 (t, 1H), 8.01 (d, 1H, J=6.8 Hz), 7.49(d, 1H, J=8 Hz), 7.33 (s, 1H), 7.18 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H,J=3.6 Hz), 3.59-3.61 (m, 6H), 2.75 (m, 1H), 2.65 (s, 3H), 2.43 (bs, 4H),2.21 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.70 (bs, 4H), 1.53-1.56 (m,1H), 1.42-1.44 (m, 1H), 1.09-1.23 (m, 4H).

Example 32: Synthesis of3-(Cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylbenzamide

To a stirred solution of compound3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(1 equiv.) and dimethylamine (5 equiv.) in methanol (10 mL), acetic acid(2 equiv.) was added and reaction stirred at room temperature for 8 h.Then sodium cyanoborohydride (2.5 equiv.) was added at 0° C. andreaction stirred overnight at room temperature. On completion, solventwas removed under reduced pressure and crude material was purified bycolumn chromatography to afford the title compound (0.017 g, 11% yield).LCMS: 516.35 (M+1)⁺; HPLC: 90.32% (@254 nm) (R_(t); 4.203; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.78(s, 1H), 8.18 (t, 1H), 8.05 (d, 1H, J=6 Hz), 7.50 (d, 1H, J=8.4 Hz),7.34 (s, 1H), 7.20 (s, 1H), 5.86 (s, 1H), 4.28 (d, 2H, J=4.8 Hz), 3.75(bs, 2H), 2.75 (m, 1H), 2.65 (s, 3H), 2.34 (bs, 6H), 2.22 (s, 3H), 2.20(s, 3H), 2.10 (s, 3H), 1.69-1.71 (m, 4H), 1.54-1.56 (m, 2H), 1.42-1.45(m, 2H), 1.08-1.23 (m, 2H).

Example 35: Synthesis of3-(Cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylbenzamide

To a stirred solution of compound3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(1 equiv.) and dimethylamine (5 equiv.) in methanol (10 mL), acetic acid(2 equiv.) was added and reaction stirred at room temperature for 18 h.Then sodium cyanoborohydride (2.5 equiv.) was added at 0° C. andreaction stirred overnight at room temperature. On completion, solventwas removed under reduced pressure and crude material was purified bycolumn chromatography to afford compound and crude material which waspurified by preparative HPLC giving the title compound as a TFA salt,(0.12 g, 57%). LCMS: 502.30 (M+1)⁺; HPLC: 99.07% (@ 254 nm) (R_(t);4.059; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.50(s, 1H), 10.04 (bs, 1H), 8.96 (s, 1H), 8.22 (m, 2H), 7.57-7.61 (m, 1H),7.35 (s, 1H), 5.87 (s, 1H), 4.49 (s, 2H), 4.28 (d, 2H, J=2 Hz), 3.65(bs, 1H), 2.83 (s, 6H), 2.65 (s, 3H), 2.28 (s, 3H), 2.12 (s, 3H), 2.10(s, 3H), 1.73 (bs, 2H), 1.63 (bs, 2H), 1.50 (m, 4H).

Example 36: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-5-(methyl(piperidin-4-yl)amino)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-(cyclopentylamino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5 g,20.57 mmol) and cyclopentanone (8.64 g, 102.8 mmol) in methanol (30 mL),acetic acid (2.46 g, 41.1 mmol) was added and reaction stirred at roomtemperature for 3 h. Then sodium cyanoborohydride (3.23 g, 51.4 mmol)was added and reaction stirred overnight. On completion, solvent wasremoved under reduced pressure and crude material was purified by columnchromatography to afford methyl5-bromo-3-(cyclopentylamino)-2-methylbenzoate (4 g, 78.2%).

Step 2: Synthesis of methyl 5-bromo-3-(cyclopentyl (ethyl)amino)-2-methylbenzoate

To a stirred solution of 5-bromo-3-(cyclopentylamino)-2-methylbenzoate(2 g, 6.43 mmol) in DMF (15 mL), cesium carbonate (4.18 g, 12.8 mmol)and ethyl iodide (5.01 g, 32.15 mmol) were added; the resulting reactionmixture was heated at 80° C. for 18 h. On completion, the reactionmixture was cooled to room temperature and filtered, residue was washedwith ethyl acetate and filtrate was concentrated to afford desired crudecompound, which was purified by column chromatography at afford methyl5-bromo-3-(cyclopentyl(ethyl)amino)-2-methylbenzoate (0.7 g, 32.1%).

Step 3: Synthesis of 5-bromo-3-(cyclopentyl (ethyl) amino)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-2-methylbenzamide

Aqueous NaOH (0.126 g, 3.09 mmol) was added to a solution of methyl5-bromo-3-(cyclopentyl (ethyl) amino)-2-methylbenzoate (0.7 g, 2.06mmol) in ethanol (5 mL) and stirred at 60° C. for 1 h. After completionof the reaction, ethanol was removed under reduced pressure and theaqueous layer acidified using dilute HCl to pH 6 and citric acid to pH4. The product was extracted using ethyl acetate. Combined organiclayers were dried and concentrated to give the crude acid (0.5 g, 75%).The acid (0.5 g, 1.53 mmol) was then dissolved in DMSO (5 mL) and3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.467 g, 3.07 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (1.19 g, 2.30 mmol) was added to it and stirring wascontinued for overnight. After completion of the reaction, the reactionmixture was poured into ice, extracted with 10% MeOH/DCM. Combinedorganic layers were dried and concentrated, then the product waspurified by column chromatography to afford 5-bromo-3-(cyclopentyl(ethyl) amino)-N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (0.3 g, 42%).

Step 4: Synthesis of 5-(cyclopentyl (ethyl) amino)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1, 1′-biphenyl]-3-carboxamide

To a stirred solution of 5-bromo-3-(cyclopentyl (ethyl) amino)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-2-methylbenzamide(0.3 g, 0.653 mmol) and (4-(morpholinomethyl) phenyl) boronic acid(0.216 g, 0.98 mmol) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (0.249g, 2.35 mmol) was added and solution purged with argon for 15 min. ThenPd (PPh₃)₄ (0.075 g, 0.065 mmol) was added and argon was purged againfor 10 min. The reaction mixture was heated at 100° C. for 3 h. Oncompletion, the reaction mixture was diluted with water and extractedwith 10% MeOH/DCM. Combined organic layers were dried over Na₂SO₄ andsolvent removed under reduced pressure to afford crude material whichwas purified by column chromatography over silica gel to afford thetitle compound (0.15 g, 41%). LCMS: 557.35 (M+1)⁺; HPLC: 99.13% (@ 254nm) (R_(t); 4.128; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.44 (s, 1H), 8.17 (t, 1H), 7.57 (d, 2H, J=8 Hz), 7.41 (s, 1H), 7.37(d, 2H, J=8 Hz), 7.20 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.8 Hz),3.56-3.57 (m, 4H), 3.48 (s, 3H), 3.00-3.02 (m, 2H), 2.36 (m, 4H), 2.24(s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.69-1.70 (m, 2H), 1.60 (m, 2H),1.47-1.48 (m, 4H), 0.81 (t, 3H, J=6.4 Hz).

Example 37: Synthesis of3-(((1r,4r)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

To a stirred solution of3-(((1r,4r)-4-acetamidocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide(1 equiv.) and4-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL) was added Na₂CO₃ (3.6equiv.) and the solution purged with argon for 15 min. Then Pd(PPh₃)₄(0.1 equiv.) was added and argon was purged again for 10 min. Thereaction mixture was heated at 100° C. for 4 h. On completion, thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic layers were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford crude material which waspurified by column chromatography over silica gel to afford the titlecompound (0.050 g, 28%). LCMS: 618.35 (M+1)⁺; HPLC: 95.34% (@ 254 nm)(R_(t); 3.760; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.47 (s, 1H), 8.17 (s, 1H), 8.09 (t, 1H), 7.82 (s, 1H), 7.67 (d, 1H,J=7.2 Hz), 7.23 (s, 1H), 7.08 (s, 1H), 5.86 (s, 1H), 4.26 (d, 2H, J=3.2Hz), 4.21 (t, 2H, J=6 Hz), 3.44-3.53 (m, 5H), 2.72 (t, 3H, J=5.6 Hz),2.61 (s, 3H), 2.40 (m, 4H), 2.20 (s, 3H), 2.13 (s, 3H), 2.10 (s, 3H),1.67-1.88 (m, 7H), 1.46-1.55 (m, 2H), 1.07-1.15 (m, 2H).

Example 38: Synthesis of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

Prepare in the analogous fashion as compound 37 (0.020 g, 11%). LCMS:618.35 (M+1)⁺; HPLC: 99.00% (@ 254 nm) (R_(t); 3.732; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.16 (s, 1H), 8.09 (t,1H), 7.82 (s, 1H), 7.77 (d, 1H, J=7.2 Hz), 7.28 (s, 1H), 7.09 (s, 1H),5.86 (s, 1H), 4.45 (bs, 1H), 4.27 (d, 2H, J=4 Hz), 4.22 (s, 2H), 3.70(bs, 1H), 3.54 (m, 4H), 2.97 (m, 1H), 2.67-2.72 (m, 2H), 2.56 (s, 3H),2.42 (m, 3H), 2.20 (s, 6H), 2.10 (s, 3H), 1.74-1.81 (m, 5H), 1.55 (m,2H), 1.39-1.41 (m, 4H).

Example 39: Synthesis of5-(((1r,4r)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of3-(((1r,4r)-4-acetamidocyclohexyl)-(methyl)-amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide(1 equiv.) andN,N-dimethyl-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanamine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL) was added Na₂CO₃ (3.6equiv.) and the solution purged with argon for 15 min. Then Pd(PPh₃)₄(0.1 equiv.) was added and argon was purged again for 10 min. Thereaction mixture was heated at 100° C. for 4 h. On completion, thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic layers were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford crude material which waspurified by column chromatography over silica gel to afford the titlecompound (0.05 g, 30%). LCMS: 572.35 (M+1)⁺; HPLC: 96.88% (@ 254 nm)(R_(t); 3.900; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 L, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.18 (t, 1H), 7.67 (d, 1H, J=6.8 Hz), 7.57 (d, 2H, J=7.6 Hz),7.34 (d, 2H, J=7.6 Hz), 7.30 (s, 1H), 7.14 (s, 1H), 5.85 (s, 1H), 4.27(d, 2H, J=3.6 Hz), 3.39 (m, 3H), 2.72 (m, 1H), 2.64 (s, 3H), 2.20 (s,6H), 2.15 (s, 6H), 2.10 (s, 3H), 1.78-1.81 (m, 2H), 1.74 (s, 3H), 1.68(m, 2H), 1.51-1.56 (m, 2H), 1.08-1.23 (m, 2H).

Example 40: Synthesis of5-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Prepared in the analogous fashion as Example 39 (0.06 g, 36%). LCMS:572.35 (M+1)⁺; HPLC: 94.79% (@ 254 nm) (R_(t); 3.936; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47 (s, 1H), 8.19 (t, 1H), 7.78 (d,1H, J=7.2 Hz), 7.56 (d, 2H, J=8 Hz), 7.33-7.35 (m, 3H), 7.17 (s, 1H),5.86 (s, 1H), 4.28 (d, 2H, J=3.6 Hz), 3.70 (bs, 1H), 3.37-3.40 (m, 2H),2.98 (m, 1H), 2.59 (s, 3H), 2.26 (s, 3H), 2.20 (m, 3H), 2.15 (s, 6H),2.10 (s, 3H), 1.81 (s, 3H), 1.74 (m, 2H), 1.55 (m, 2H), 1.40-1.48 (m,4H).

Example 41: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methyl-3-(methyl(piperidin-4-yl)amino)benzamide

To a stirred solution of tert-butyl4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)piperidine-1-carboxylate(1 equiv.) and morpholine (5 equiv.) in methanol (5 mL for 0.3 mmol),acetic acid (1 equiv.) was added and reaction stirred at roomtemperature for 4 h. Then reducing agent NaBH₃CN (1 equiv.) was addedand reaction stirred overnight. On completion, solvent was removed underreduced pressure and residue purified by column chromatography oversilica gel affording desired tert-butyl4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylphenyl)(methyl)amino)piperidine-1-carboxylate.This compound was then dissolved in DCM (5 mL) and cooled to 0° C. TFA(2 mL) was added to it. The reaction mixture was stirred at roomtemperature for 1 h. On completion, reaction was concentrated todryness. Residue was purified by solvent washings to afford the titlecompound (0.06 g, 40%). LCMS: 517.25 (M+1)⁺; HPLC: 99.07% (@ 254 nm)(R_(t); 3.913; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.48 (s, 1H), 10.08 (bs, 1H), 8.97 (s, 1H), 8.57 (bs, 1H), 8.23 (d, 2H,J=7.6 Hz), 8.18 (s, 1H), 7.60 (d, 1H, J=8 Hz), 7.50 (s, 1H), 7.34 (s,1H), 5.87 (s, 1H), 4.49 (d, 2H), 4.30 (s, 2H), 3.25 (d, 2H), 3.16 (s,1H), 2.89 (m, 2H), 2.83 (s, 6H), 2.64 (s, 3H), 2.26 (s, 3H), 2.21 (s,3H), 2.10 (s, 3H), 1.81 (bs, 4H).

Example 42: Synthesis of5-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(methyl)-amino)-2-methylbenzoate

To a stirred solution of the less polar cis isomer, methyl5-bromo-3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate,(4 g, 9.09 mmol) in acetonitrile (50 mL), cesium carbonate (5.9 g, 18.18mmol) and methyl iodide (6.45 g, 45.45 mmol) were added. The resultingreaction mixture was heated at 80° C. for 7 h. The reaction mixture wascooled to room temperature and filtered, with the collected solids beingwashed with ethyl acetate. The filtrate was concentrated to afforddesired product which purified by column chromatography giving methyl5-bromo-3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(methyl)-amino)-2-methylbenzoate(1.4 g, 34.14%).

Step 2: Synthesis of tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate

Aqueous NaOH (0.23 g, 5.72 mmol) was added to a solution of methyl5-bromo-3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(methyl)-amino)-2-methylbenzoate(1.3 g, 2.86 mmol) in MeOH (20 mL) and stirred at 60° C. for 1 h. Theethanol was removed under reduced pressure and the mixture acidified topH with dilute HCl and to pH 4 with citric acid. The mixture wasextracted with ethyl acetate. The combined organic extracts were driedand concentrated giving respective acid (1.13 g, 90.1%).

The acid (1.13 g, 2.57 mmol) was then dissolved in DMSO (10 mL) and3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.87 g, 5.72 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PyBOP (2.23 g, 4.28 mmol) was added. Stirring was thencontinued overnight. The reaction, reaction mixture was poured into icewater. The resulting precipitate was filtered, washed with acetonitrileand purified by column chromatography to afford tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate(0.8 g, 48.7%).

Step 3: Synthesis of tert-butyl((1s,4s)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)-cyclohexyl)carbamate

To a stirred solution of tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)-carbamate(1 equiv.) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine (1.2equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) wasadded. The solution was then purged with argon for 15 min. Pd(PPh₃)₄(0.1 equiv.) was added and the reaction mixture again purged with argonfor 10 min. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined extracts were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford the crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl((1s,4s)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)-cyclohexyl)carbamate(0.08 g, 45.71%).

Step 4: Synthesis of5-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A stirred solution of tert-butyl((1s,4s)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)-cyclohexyl)carbamate(0.08 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness yielding the title compound as a TFA salt(0.06 g, 88.2%). LCMS: 572.40 (M+1)⁺; HPLC: 95.39% (@ 254 nm) (R_(t);3.719; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 10.05 (bs, 1H), 8.19 (t, 1H), 7.74-7.78 (m, 4H), 7.56 (d, 2H,J=6.8 Hz), 7.46 (s, 1H), 7.24 (s, 1H), 5.87 (s, 1H), 4.38 (bs, 2H), 4.29(d, 2H, J=4.4 Hz), 3.95 (m, 2H), 3.60-3.63 (m, 2H), 3.27-3.30 (m, 2H),3.13-3.19 (m, 4H), 2.54 (s, 3H), 2.30 (s, 3H), 2.21 (s, 3H), 2.10 (s,3H), 1.86 (m, 2H), 1.59-1.64 (m, 4H), 1.49-1.51 (m, 2H).

Example 43: Synthesis of5-(((1r,4r)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic acid

To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g, 276.2 mmol)in conc. H₂SO₄ (200 mL), 1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione(43.4 g, 151.8 mmol) was added portionwise at room temperature and thereaction mixture stirred at room temperature for 5 h. The reactionmixture was poured into ice cold water; the solid which precipitated wasfiltered, washed with water and dried under vacuum giving the desiredcompound, 5-bromo-2-methyl-3-nitrobenzoic acid (71.7 g, 99.9%) which wasused as is in further reactions.

Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate

To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid (287 g,1103 mmol) in DMF (150 mL), sodium carbonate (468 g, 4415 mmol) andmethyl iodide (626.63 g, 4415 mmol) were added. The reaction mixture wasthen heated at 60° C. for 8 h. The precipitated solids were filtered andwashed with diethyl ether (5 times). The combined organic filtrates weredried, concentrated under reduced pressure giving desired compoundmethyl 5-bromo-2-methyl-3-nitrobenzoate (302 g, 99%) which was used asis in further reactions

Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate

To a stirred solution of methyl 5-bromo-2-methyl-3-nitrobenzoate (150 g,544 mmol) in ethanol (750 mL), ammonium chloride (150 g, 2777 mmol)dissolved in water (750 mL) and iron powder (93.3 g, 1636 mmol) wereadded under stirring. The resulting reaction mixture was heated at 80°C. for 7 h. The reaction mixture was filtered through Celite and thecollected solids washed with water and ethyl acetate. The filtrate wasextracted with ethyl acetate and the extract dried, concentrated underreduced pressure to give the desired compound methyl3-amino-5-bromo-2-methylbenzoate which was used as is in furtherreactions.

Step 4: Synthesis of methyl5-bromo-3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methyl-benzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5.0 g,20.6 mmol) and tert-butyl (4-oxocyclohexyl)carbamate (5.6 g, 26.7 mmol)in methanol (50 mL), acetic acid (1.2 g, 20.57 mmol) was added andreaction mixture stirred at room temperature for 8 h. Then sodiumcyanoborohydride (1.6 g, 26.74 mmol) was added at 0° C. and the reactionstirred overnight. The solvent was removed under reduced pressure andthe crude material purified by column chromatography (twice) elutingwith ethyl acetate/hexane to afford 4 g (44%) of less-polar cis isomer,methyl5-bromo-3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate(contaminated with some starting material) and 3 g (33%) of the morepolar pure trans isomer, methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate.

Step 5: Synthesis of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(methyl)-amino)-2-methylbenzoate

To a stirred solution of the more polar trans isomer, methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate,(3 g, 6.81 mmol) in acetonitrile (40 mL), cesium carbonate (4.4 g, 13.62mmol) and methyl iodide (4.83 g, 34.05 mmol) were added The resultingreaction mixture was heated at 80° C. for 7 h. The reaction mixture wascooled to room temperature and filtered and the solids washed with ethylacetate. The filtrate was concentrated to afford the desired crudecompound which purified by column chromatography giving methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(methyl)amino)-2-methylbenzoate(1.3 g, 43.33%).

Step 6: Synthesis of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate

Aqueous NaOH (0.23 g, 5.72 mmol) was added to a solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(methyl)amino)-2-methylbenzoate(1.3 g, 2.86 mmol) in MeOH (20 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, the methanol was removed under reducedpressure and the residue acidified to pH 6 with dilute HCl and to pH 4with citric acid. The acidified mixture was extracted with ethylacetate. The combined organic extracts were dried and concentratedgiving the respective acid (1 g, 83%).

The above acid (1 g, 2.27 mmol) was dissolved in DMSO (5 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.65 g, 4.54 mmol) wasadded. The reaction mixture was stirred at room temperature for 15 minbefore PyBOP (1.7 g, 3.4 mmol) was added. Stirring was continuedovernight. The reaction mixture was poured into ice water. The resultingprecipitate was filtered, washed with acetonitrile and purified bycolumn chromatography to afford compound tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(0.7 g, 53.8%).

Step 7: Synthesis of tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-4-methy-4′-(mrpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)-cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(1 equiv.) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine (1.2equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) wasadded and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and the reaction flask was purged again for 10 min.with argon. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by column chromatography over silica gel to afford tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate(0.07 g, 40%)

Step 8: Synthesis of5-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A stirred solution of tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate(0.07 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness yielding the title compound as a TFA salt(0.05 g, 84.74%). LCMS: 572.60 (M+1)⁺; HPLC: 88.92% (@ 254 nm) (R_(t);3.546; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 10.05 (bs, 1H), 8.16 (t, 1H), 7.74-7.76 (m, 4H), 7.56 (d, 2H,J=7.6 Hz), 7.34 (s, 1H), 7.21 (s, 1H), 5.86 (s, 1H), 4.38 (bs, 2H), 4.28(d, 2H, J=4.4 Hz), 3.95 (m, 2H), 3.63 (m, 2H), 3.27 (m, 1H), 3.12 (m,2H), 2.97 (m, 2H), 2.74 (t, 1H), 2.66 (s, 3H), 2.20 (s, 6H), 2.10 (s,3H), 1.93-1.95 (m, 2H), 1.74-1.77 (m, 2H), 1.54-1.57 (m, 2H), 1.28-1.31(m, 2H).

Example 44: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic acid

To stirred solution of 2-methyl-3-nitrobenzoic acid (100 g, 552 mmol) inconc. H₂SO₄ (400 mL), 1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione(88 g, 308 mmol) was added in a portion wise manner at room temperatureand the reaction mixture was then stirred at room temperature for 5 h.The reaction mixture was poured onto ice cold water, the precipitatedsolid was filtered off, washed with water and dried under vacuum toafford the desired compound as a solid (140 g, 98%). The isolatedcompound was taken directly into the next step. ¹H NMR (DMSO-d₆, 400MHz) δ 8.31 (s, 1H), 8.17 (s, 1H), 2.43 (s, 3H).

Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate

To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid (285 g,1105 mmol) in DMF (2.8 L) at room temperature was added sodium carbonate(468 g, 4415 mmol) followed by addition of methyl iodide (626.6 g, 4415mmol). The resulting reaction mixture was heated at 60° C. for 8 h.After completion (monitored by TLC), the reaction mixture was filtered(to remove sodium carbonate) and washed with ethyl acetate (1 L×3). Thecombined filtrate was washed with water (3 L×5) and the aqueous phasewas back extracted with ethyl acetate (1 L×3). The combined organiclayers were dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure to afford the title compound as asolid (290 g, 97% yield). The isolated compound was taken directly intothe next step. ¹H NMR (CDCl₃, 400 MHz) δ 8.17 (s, 1H), 7.91 (s, 1H),3.96 (s, 3H), 2.59 (s, 3H).

Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate

To a stirred solution of methyl 5-bromo-2-methyl-3-nitrobenzoate (290 g,1058 mmol) in ethanol (1.5 L) was added aqueous ammonium chloride (283g, 5290 mmol dissolved in 1.5 L water). The resulting mixture wasstirred at 80° C. to which iron powder (472 g, 8451 mmol) was added in aportion wise manner. The resulting reaction mixture was heated at 80° C.for 12 h. Upon completion as determined by TLC, the reaction mixture washot filtered over Celite® and the celite bed was washed with methanol (5L) followed by washing with 30% MeOH in DCM (5 L). The combined filtratewas concentrated in-vacuo, the residue obtained was diluted with aqueoussodium bicarbonate solution (2 L) and extracted with ethyl acetate (5L×3). The combined organic layers were dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure to afford thetitle compound as a solid (220 g, 85%). The compound was taken directlyinto the next step. ¹H NMR (CDCl₃, 400 MHz) δ 7.37 (s, 1H), 6.92 (s,1H), 3.94 (s, 3H), 3.80 (bs, 2H), 2.31 (s, 3H).

Step 4: Synthesis of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (15 g,61.5 mmol) and dihydro-2H-pyran-4(3)-one (9.2 g, 92 mmol) indichloroethane (300 mL) was added acetic acid (22 g, 369 mmol) and thereaction mixture stirred at room temperature for 15 minutes, then thereaction mixture was cooled to 0° C. and sodium triacetoxyborohydride(39 g, 184 mmol) was added. The reaction mixture was stirred overnightat room temperature. Upon completion of the reaction as determined byTLC, aqueous sodium bicarbonate solution was added to the reactionmixture until a pH of 7-8 was obtained. The organic phase was separatedand the aqueous phase was extracted with ethyl acetate. The combinedorganic layers were dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure. The crude compound was purified bycolumn chromatography (100-200 mesh silica gel) eluting with ethylacetate: hexane to afford the desired compound as a solid (14 g, 69%).¹H NMR (DMSO-d₆, 400 MHz) δ 7.01 (s, 1H), 6.98 (s, 1H), 5.00 (d, 1H,J=7.6 Hz), 3.84-3.87 (m, 2H), 3.79 (s, 3H), 3.54-3.56 (m, 1H), 3.43 (t,2H, J=12 Hz), 2.14 (s, 3H), 1.81-1.84 (m, 2H), 1.47-1.55 (m, 2H).

Step 5: Synthesis of methyl 5-bromo-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate (14 g,42.7 mmol) in dichloroethane (150 mL) was added acetaldehyde (3.75 g,85.2 mmol) and acetic acid (15.3 g, 256 mmol). The resulting reactionmixture was stirred at room temperature for 15 minutes. The mixture wascooled to 0° C. and sodium triacetoxyborohydride (27 g, 128 mmol) wasadded. The reaction mixture was stirred at room temperature for 3 hours.Upon completion of the reaction as determined by TLC, aqueous sodiumbicarbonate solution was added to the reaction mixture until a pH 7-8was obtained, the organic phase was separated and the aqueous phase wasextracted with ethyl acetate. The combined organic layers were driedover anhydrous sodium sulfate, filtered and concentrated under reducedpressure. The crude compound was purified by column chromatography(100-200 mesh silica gel) eluting with ethyl acetate: hexane to affordthe desired compound as a viscous liquid (14 g, 93%). ¹H NMR (DMSO-d₆,400 MHz) δ 7.62 (s, 1H), 7.52 (s, 1H), 3.80 (bs, 5H), 3.31 (t, 2H),2.97-3.05 (m, 2H), 2.87-2.96 (m, 1H), 2.38 (s, 3H), 1.52-1.61 (m, 2H),1.37-1.50 (m, 2H), 0.87 (t, 3H, J=6.8 Hz).

Step 6: Synthesis of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

To a stirred solution of 5-bromo-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate (14 g, 39.4 mmol) in ethanol (100 mL) was addedaqueous NaOH (2.36 g, 59.2 mmol in 25 mL water) and the resultingmixture was stirred at 60° C. for 1 h. Upon completion of the reactionas determined by TLC, the solvent was removed under reduced pressure andthe residue obtained was acidified with 1N HCl until a pH 7 was obtainedand then aqueous citric acid solution was added until a pH 5-6 wasobtained. The aqueous layer was extracted with 10% MeOH in DCM (200mL×3), the combined organic layers were dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure to give therespective acid (14 g, 100%).

The above acid (14 g, 40.9 mmol) was then dissolved in DMSO (70 mL) and3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (12.4 g, 81.9 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15minutes, then PYBOP (31.9 g, 61.4 mmol) was added and stirring wascontinued for overnight at room temperature. Upon completion of thereaction as determined by TLC, the reaction mixture was poured ontoice-cold water (700 mL), stirred for 30 minutes and the precipitatedsolid was collected by filtration, washed with water (500 mL) and airdried. The solid obtained was stirred with acetonitrile (75 mL×2),filtered and air dried. The solid obtained was again stirred with 5%MeOH in DCM (100 mL), filtered and dried completely under vacuum toafford the title compound as a solid (14 g, 74%). ¹H NMR (DMSO-d₆, 400MHz) δ 11.47 (s, 1H), 8.23 (t, 1H), 7.30 (s, 1H), 7.08 (s, 1H), 5.85 (s,1H), 4.23 (d, 2H, J=4.4 Hz), 3.81 (d, 2H, J=10.4 Hz), 3.20-3.26 (m, 2H),3.00-3.07 (m, 1H), 2.91-2.96 (m, 2H), 2.18 (s, 3H), 2.14 (s, 3H), 2.10(s, 3H), 1.58-1.60 (m, 2H), 1.45-1.50 (m, 2H), 0.78 (t, 3H, J=6.8 Hz).

Step 7: Synthesis of N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1, 1′-biphenyl]-3-carboxamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (14 g, 29.5 mmol) in dioxane/water mixture (70mL/14 mL) was added 4-(4-(4, 4, 5, 5-tetramethyl-1, 3,2-dioxaborolan-2-yl) benzyl) morpholine (13.4 g, 44.2 mmol) followed byaddition of Na₂CO₃ (11.2 g, 106.1 mmol). The solution was purged withargon for 15 minutes and then Pd (PPh₃)₄ (3.40 g, 2.94 mmol) was addedand the solution was again purged with argon for a further 10 min. Thereaction mixture was heated at 100° C. for 4 h. After completion(monitored by TLC), the reaction mixture was diluted with water andextracted with 10% MeOH/DCM. The combined organic layers were dried overanhydrous sodium sulphate, filtered and concentrated under reducedpressure. The crude compound was purified by column chromatography(100-200 mesh silica gel) eluting with methanol: DCM to the titlecompound as a solid (12 g, 71%). Analytical Data: LCMS: 573.35 (M+1)⁺;HPLC: 99.5% (@ 254 nm) (R_(t); 3.999; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 11.46 (s, 1H), 8.19 (t, 1H), 7.57 (d,2H, J=7.2 Hz), 7.36-7.39 (m, 3H), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d,2H, J=2.8 Hz), 3.82 (d, 2H, J=9.6 Hz), 3.57 (bs, 4H), 3.48 (s, 2H), 3.24(t, 2H, J=10.8 Hz), 3.07-3.09 (m, 2H), 3.01 (m, 1H), 2.36 (m, 4H), 2.24(s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m, 2H), 1.51-1.53 (m,2H), 0.83 (t, 3H, J=6.4 Hz).

Step 8: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamidetrihydrochloride

N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide (12 g, 21.0 mmol) was dissolved in methanolicHCl (200 mL) and stirred at room temperature for 3 h. After three hoursof stirring, the reaction mixture was concentrated under reducedpressure. The solid obtained was stirred with ether (100 mL×2) to affordthe desired salt as a solid (11 g, 77%). Analytical Data of the tri-HClsalt: LCMS: 573.40 (M+1)⁺; HPLC: 99.1% (@ 254 nm) (R_(t); 3.961; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (D₂O 400 MHz) δ 7.92 (bs, 1H) 7.80 (s,1H), 7.77 (d, 2H, J=8 Hz), 7.63 (s, 1H), 7.61 (s, 1H), 6.30 (s, 1H),4.48 (s, 2H), 4.42 (s, 2H), 4.09-4.11 (m, 4H), 3.95-3.97 (m, 2H), 3.77(t, 3H, J=10.4 Hz), 3.44-3.47 (m, 3H), 3.24-3.32 (m, 3H), 2.42 (s, 3H),2.35 (s, 3H), 2.26 (s, 3H), 2.01 (m, 2H), 1.76 (m, 2H), 1.04 (t, 3H,J=6.8 Hz).

Example 45: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

Step 1: Synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(1 equiv.) and1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and the reaction flask was purged again for 10 min.with argon. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by column chromatography over silica gel to afford tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.07 g, 46.6%)

Step 2: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

A stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.07 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness yielding the title compound as a TFA salt(0.07 g, 98.59%). LCMS: 477.35 (M+1)⁺; HPLC: 99.16% (@ 254 nm) (R_(t);3.796; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.12 (s, 1H), 8.08 (t, 1H), 7.82 (s, 1H), 7.74 (m, 3H), 7.28(s, 1H), 7.11 (s, 1H), 5.86 (s, 1H), 4.26 (d, 2H, J=4.4 Hz), 3.84 (s,3H), 2.96 (bs, 1H), 2.73 (bs, 1H), 2.63 (s, 3H), 2.20 (s, 3H), 2.14 (s,3H), 2.10 (s, 3H), 1.92-1.95 (m, 2H), 1.74-1.77 (m, 2H), 1.48-1.57 (m,2H), 1.23-1.32 (m, 2H).

Example 46: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

Step 1: Synthesis of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)-carbamate(1 equiv.) and1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added. The solution was then purged with argon for 15 min. Pd(PPh₃)₄(0.1 equiv.) was added and the reaction mixture again purged with argonfor 10 min. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined extracts were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford the crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.05 g, 33.3%).

Step 2: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

A stirred solution of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.05 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness yielding the title compound as a TFA salt(0.03 g, 73.1%). LCMS: 477.30 (M+1)⁺; HPLC: 98.76% (@ 254 nm) (R_(t);3.862; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.08-8.12 (m, 2H), 7.76-7.81 (m, 4H), 7.33 (s, 1H), 7.12 (s,1H), 5.86 (s, 1H), 4.27 (d, 2H, J=4 Hz), 3.83 (s, 3H), 3.16 (m, 2H),2.50 (3H merged in solvent peak), 2.22 (s, 3H), 2.20 (s, 3H), 2.10 (s,3H), 1.84 (m, 2H), 1.57-1.63 (m, 4H), 1.47-1.50 (m, 2H).

Example 47: Synthesis of5-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of tert-butyl((1s,4s)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)-carbamate(1 equiv.) andN,N-dimethyl-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanamine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added. The solution was purged then with argon for 15 min. Pd(PPh₃)₄(0.1 equiv.) was added and the reaction mixture again purged with argonfor 10 min. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined extracts were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford the crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl((1s,4s)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate(0.100 g, 61%).

Step 2: Synthesis of5-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

A stirred solution of tert-butyl((1s,4s)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate(0.10 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness yielding the title compound as a TFA salt(0.05 g, 59.5%). LCMS: 530.35 (M+1)⁺; HPLC: 97.13% (@ 254 nm) (R_(t);3.672; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.44(s, 1H), 9.47 (bs, 1H), 8.17 (t, 1H), 7.74-7.76 (m, 4H), 7.55 (d, 2H,J=7.6 Hz), 7.44 (s, 1H), 7.25 (s, 1H), 5.86 (s, 1H), 4.30 (m, 4H), 3.12(m, 2H), 2.74 (s, 6H), 2.54 (s, 3H), 2.30 (s, 3H), 2.12 (s, 3H), 2.10(s, 3H), 1.84 (bs, 2H), 1.59-1.63 (m, 4H), 1.48 (m, 2H).

Example 48: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

Step 1: Synthesis of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)-carbamate(1 equiv.) and4-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added. The solution was purged then with argon for 15 min. Pd(PPh₃)₄(0.1 equiv.) was added and the reaction mixture again purged with argonfor 10 min. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined extracts were dried over Na₂SO₄ and the solvent removedunder reduced pressure to afford the crude product which was purified bycolumn chromatography over silica gel to afford tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.120 g, 75.4%).

Step 2: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

A stirred solution of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.10 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness yielding the title compound as a TFA salt(0.06 g, 58.82%). LCMS: 576.40 (M+1)⁺; HPLC: 96.89% (@ 254 nm) (R_(t);3.481; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.25 (s, 1H), 8.08 (t, 1H), 7.79 (s, 1H), 7.74-7.79 (m, 3H),7.34 (s, 1H), 7.15 (s, 1H), 5.86 (s, 1H), 4.51 (bs, 2H), 4.27 (d, 2H,J=4.4 Hz), 3.16 (m, 6H), 2.50 (3H merged in solvent peak), 2.23 (s, 3H),2.21 (s, 3H), 2.11 (s, 3H), 1.84 (bs, 2H), 1.57-1.63 (m, 4H), 1.47-1.49(m, 2H). [3H merged in solvent peak].

Example 49: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Synthesis of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)-amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(0.5 g, 8.71 mmol) and (6-formylpyridin-3-yl) boronic acid (0.264 g,1.13 mmol) in dioxane/water mixture (10 mL+2 mL), Na₂CO₃ (0.333 g, 2.8mmol) was added. The solution was then purged with argon for 15 min. Pd(PPh₃)₄ (0.1 g, 0.086 mmol) was added and the solution again purged withargon for 10 min. The reaction mixture was heated at 100° C. for 4 h.The reaction mixture was diluted with water and extracted with 10%MeOH/DCM. The combined extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by column chromatography over silica gel to afford tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate(0.3 g, 57.3%).

Step 2: Synthesis of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)phenyl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate(1 equiv.) and morpholine (5 equiv.) in methanol (10 mL), acetic acid (2equiv.) was added. The reaction mixture was stirred at room temperaturefor 18 h. Sodium cyanoborohydride (2.5 equiv.) was then added at 0° C.and the reaction mixture stirred overnight at room temperature. Thesolvent was removed under reduced pressure and the crude product waspurified by column chromatography to afford tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)phenyl)(methyl)amino)cyclohexyl)carbamate.

Step 3: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

A stirred solution of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)phenyl)(methyl)amino)cyclohexyl)carbamatein DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. The reactionmixture was stirred at room temperature for 1 h. The reaction wasconcentrated to dryness and the product purified by solvent washings toafford the title compound as a TFA salt (0.1 g, 94.33%). LCMS: 573.45(M+1)⁺; HPLC: 98.94% (@ 254 nm) (R_(t); 3.618; Method: Column: YMC ODS-A150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47 (s, 1H), 8.94 (s, 1H),8.19-8.21 (m, 2H), 7.80 (s, 3H), 7.60 (d, 1H, J=8 Hz), 7.49 (s, 1H),7.31 (s, 1H), 5.86 (s, 1H), 4.52 (bs, 2H), 4.29 (d, 2H, J=4.4 Hz), 3.83(bs, 4H), 3.27 (m, 4H), 3.14-3.21 (m, 2H), 2.55 (s, 3H), 2.30 (s, 3H),2.21 (s, 3H), 2.10 (s, 3H), 1.87 (bs, 2H), 1.59-1.64 (m, 4H), 1.49-1.51(m, 2H).

Example 50: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)cyclohexyl)-carbamate(0.4 g, 0.696 mmol) and (6-formylpyridin-3-yl)boronic acid (0.21 g,0.906 mmol) in dioxane/water mixture (8 mL+2 mL), Na₂CO₃ (0.332 g, 3.13mmol) was added. The reaction solution was then purged with argon for 15min. Pd (PPh₃)₄ (0.080 g, 0.069 mmol) was added and argon purging wasagain performed for 10 min. The reaction mixture was heated at 100° C.for 4 h. The reaction mixture was diluted with water and extracted with10% MeOH/DCM. The combined organic layers were dried over Na₂SO₄ and thesolvent removed under reduced pressure to afford crude product which waspurified by column chromatography over silica gel to afford tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(0.28 g, 66.98%).

Step 2: Synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)phenyl)(methyl)-amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(1 equiv.) and morpholine (5 equiv.) in methanol (10 mL), acetic acid (2equiv.) was added. The reaction was stirred at room temperature for 18h. Sodium cyanoborohydride (2.5 equiv.) was then added at 0° C. andreaction stirred overnight at room temperature. The solvent was removedunder reduced pressure and crude material was purified by columnchromatography to afford tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)phenyl)(methyl)-amino)cyclohexyl)carbamate.

Step 3: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

A stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)phenyl)(methyl)-amino)cyclohexyl)carbamatein DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added to it.Reaction mass was stirred at room temperature for 1 h. The reactionmixture was concentrated to dryness and the solid product purified bysolvent washings to afford the title compound as a TFA salt (0.07 g,82.3%). LCMS: 573.40 (M+1)⁺; HPLC: 91.56% (@ 254 nm) (R_(t); 3.591;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H),8.95 (s, 1H), 8.19-8.22 (m, 2H), 7.78 (bs, 3H), 7.61 (d, 1H, J=8 Hz),7.40 (s, 1H), 7.27 (s, 1H), 5.86 (s, 1H), 4.52 (bs, 2H), 4.28 (d, 2H,J=3.2 Hz), 3.84 (bs, 4H), 3.27 (bs, 4H), 2.97 (bs, 1H), 2.75 (m, 1H),2.66 (s, 3H), 2.21 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.93 (m, 2H),1.74-1.76 (m, 2H), 1.54-1.57 (m, 2H), 1.28-1.31 (m, 2H).

Example 51: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

Step 1: Synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(1 equiv.) and4-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethyl)morpholine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and the reaction flask was purged again for 10 min.with argon. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by column chromatography over silica gel to afford tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.08 g, 45.45%)

Step 2: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)benzamide

A stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)phenyl)(methyl)amino)cyclohexyl)carbamate(0.08 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness the title compound as a TFA salt (0.07 g,86.41%). LCMS: 576.45 (M+1)⁺; HPLC: 98.26% (@ 254 nm) (R_(t); 3.413;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H),8.26 (s, 1H), 8.08 (t, 1H), 7.99 (s, 1H), 7.75 (m, 3H), 7.28 (s, 1H),7.13 (s, 1H), 5.87 (s, 1H), 4.53 (t, 2H), 4.27 (d, 2H, J=3.6 Hz),2.97-3.16 (m, 4H), 2.67-2.71 (m, 1H), 2.62 (s, 3H), 2.20 (s, 3H), 2.14(s, 3H), 2.11 (s, 3H), 1.92-1.94 (m, 2H), 1.72 (m, 2H), 1.52-1.55 (m,2H), 1.23-1.29 (m, 2H).

Example 52: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylbenzamide

Step 1: Synthesis of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-carbamoyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate

To a stirred solution of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)-(methyl)amino)cyclohexyl)carbamate(1 equiv.) and dimethylamine (5 equiv.) in methanol (10 mL), acetic acid(2 equiv.) was added. The reaction mixture was stirred at roomtemperature for 18 h. Sodium cyanoborohydride (2.5 equiv.) was thenadded at 0° C. and the reaction mixture stirred overnight at roomtemperature. The solvent was removed under reduced pressure and thecrude product was purified by column chromatography to afford tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate.

Step 2: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylbenzamide

A stirred solution of tert-butyl((1s,4s)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamatein DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. The reactionmixture was stirred at room temperature for 1 h. The reaction wasconcentrated to dryness and the product purified by solvent washings tothe title compound as a TFA salt (0.07 g, 93.3%). LCMS: 531.25 (M+1)⁺;HPLC: 97.59% (@ 254 nm) (R_(t); 3.680; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 10.01 (s, 1H), 8.95(s, 1H), 8.20 (d, 2H, J=5.2 Hz), 7.80 (bs, 3H), 7.59 (d, 1H, J=8 Hz),7.51 (s, 1H), 7.32 (s, 1H), 5.87 (s, 1H), 4.48 (bs, 2H), 4.29 (d, 2H,J=4.4 Hz), 3.21 (m, 1H), 3.14-3.16 (m, 1H), 2.83 (s, 6H), 2.55 (s, 3H),2.31 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.86 (bs, 2H), 1.59-1.64 (m,4H), 1.49-1.51 (m, 2H).

Example 53: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylbenzamide

Step 1: Synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-formylpyridin-3-yl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(1 equiv.) and dimethylamine (5 equiv.) in methanol (10 mL), acetic acid(2 equiv.) was added. The reaction was stirred at room temperature for18 h. Sodium cyanoborohydride (2.5 equiv.) was then added at 0° C. andreaction stirred overnight at room temperature. The solvent was removedunder reduced pressure and crude material was purified by columnchromatography to afford tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate.

Step 2: Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylbenzamide

A stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamatein DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added to it.Reaction mass was stirred at room temperature for 1 h. The reactionmixture was concentrated to dryness and the solid product purified bysolvent washings to afford the title compound as a TFA salt (0.05 g,66.6%). LCMS: 531.30 (M+1)⁺; HPLC: 97.59% (@ 254 nm) (R_(t); 3.564;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H),10.01 (s, 1H), 8.95 (s, 1H), 8.20 (bs, 2H), 7.78 (bs, 2H), 7.59 (d, 1H,J=6 Hz), 7.41 (s, 1H), 7.28 (s, 1H), 5.86 (s, 1H), 4.48 (bs, 2H), 4.29(m, 2H), 2.97 (bs, 2H), 2.83 (s, 6H), 2.66 (s, 3H), 2.21 (s, 6H), 2.10(s, 3H), 1.93 (m, 2H), 1.74 (m, 2H), 1.55-1.57 (m, 2H), 1.28-1.31 (m,2H).

Example 54: Synthesis of3-(((1r,4r)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Compound 54 was prepared with the method similar to that described inExample 57.

Analytical Data of: LCMS: 615.55 (M+1)⁺; HPLC: 98.75% (@ 254 nm) (R_(t);3.854; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.75 (s, 1H), 8.18 (t, 1H), 8.02 (d, 1H, J=8 Hz), 7.67 (d, 1H,J=7.2 Hz), 7.49 (d, 1H, J=8 Hz), 7.35 (s, 1H), 7.19 (s, 1H), 5.86 (s,1H), 4.28 (d, 2H, J=4.4 Hz), 3.59-3.61 (m, 4H), 3.47-3.55 (m, 2H), 2.76(t, 2H, J=4 Hz), 2.65 (s, 3H), 2.42 (bs, 4H), 2.21 (s, 3H), 2.20 (s,3H), 2.10 (s, 3H), 1.78-1.90 (m, 2H), 1.68-1.74 (m, 5H), 1.48-1.57 (m,2H), 1.03-1.23 (m, 2H).

Example 55: Synthesis of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-2-methylbenzamide

Step 1: Synthesis of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide

3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.65 g, 1.25 mmol) and (6-formylpyridin-3-yl)boronic acid (0.38 g, 1.63mmol) in dioxane/water mixture (10 mL+2 mL) was added Na₂CO₃ (0.48 g,4.53 mmol) and the solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.14 g, 0.12 mmol) was added and argon was purged again for 10min. The reaction mixture was heated at 100° C. for 4 h. On completion,the reaction mixture was diluted with water and extracted with 10%MeOH/DCM. The combined organic layers were dried over Na₂SO₄ and solventremoved under reduced pressure to afford crude material which waspurified by column chromatography over silica gel to afford cis-isomer3-((4-acetamidocyclohexyl)-(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(0.35 g, 51.16%).

Step 2: Synthesis of To a stirred solution of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(1 equiv.) and dimethylamine (5 equiv.) in 5 mL for 0.3 mmol; MeOH wasadded acetic acid (2 equiv.) and the reaction stirred at roomtemperature. Then NaBH₃CN (1.5 equiv.) was added and the reactionstirred overnight. On completion, the solvent was removed under reducedpressure and the residue purified by column chromatography over silicagel or as specified affording the title compound (0.006 g, 3.2%). LCMS:573.40 (M+1)⁺; HPLC: 95.52% (@ 254 nm) (R_(t); 3.899; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47 (s, 1H), 8.88 (s, 1H), 8.20 (t,1H), 8.14 (d, 1H, J=7.6 Hz), 7.78 (d, 1H, J=7.2 Hz), 7.55 (d, 1H, J=8Hz), 7.44 (s, 1H), 7.26 (s, 1H), 5.86 (s, 1H), 4.28 (d, 2H, J=3.2 Hz),4.26 (bs, 1H), 3.71 (bs, 1H), 3.01 (bs, 1H), 2.61-2.66 (m, 8H), 2.28 (s,3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.81 (m, 5H), 1.56 (m, 2H), 1.40-1.46(m, 2H), 1.23 (m, 2H). [2H merged in solvent peak].

Example 56: Synthesis of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-(hydroxymethyl)pyridin-3-yl)-2-methylbenzamide

Compound 56 was prepared in the same reaction as compound 55. LCMS:546.40 (M+1)⁺; HPLC: 99.40% (@ 254 nm) (R_(t); 3.845; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47 (s, 1H), 8.74 (s, 1H), 8.20 (t,1H), 8.04 (d, 1H, J=8 Hz), 7.77 (d, 1H, J=7.2 Hz), 7.52 (d, 1H, J=7.6Hz), 7.40 (s, 1H), 7.22 (s, 1H), 5.86 (s, 1H), 5.45 (t, 1H, J=5.2 Hz),4.59 (d, 2H, J=5.6 Hz), 4.27 (d, 2H, J=4 Hz), 3.71 (bs, 1H), 3.00 (bs,1H), 2.60 (s, 3H), 2.27 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.81 (m,5H), 1.56 (m, 2H), 1.40-1.48 (m, 4H).

Example 57: Synthesis of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

To a stirred solution of3-(((1s,4s)-4-acetamidocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(1 equiv.) and morpholine (5 equiv.) in 5 mL for 0.3 mmol; MeOH wasadded acetic acid (2 equiv.) and the reaction stirred at roomtemperature. Then NaBH₃CN (1.5 equiv.) was added and the reactionstirred overnight. On completion, the solvent was removed under reducedpressure and the residue purified by column chromatography over silicagel or as specified affording the title compound (0.08 g, 43%). LCMS:615.40 (M+1)⁺; HPLC: 99.64% (@ 254 nm) (R_(t); 3.900; Method: Column:YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05%TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.75 (s, 1H), 8.19 (t,1H), 8.01 (d, 1H, J=7.6 Hz), 7.77 (d, 1H, J=7.2 Hz), 7.50 (d, 1H, J=8Hz), 7.40 (s, 1H), 7.21 (s, 1H), 5.86 (s, 1H), 4.28 (d, 2H, J=4.4 Hz),3.71 (bs, 1H), 3.59-3.61 (m, 4H), 3.50 (t, 1H, J=4.4 Hz), 3.00 (bs, 1H),2.68 (t, 1H, J=4.4 Hz), 2.60 (s, 3H), 2.42 (bs, 4H), 2.27 (s, 3H), 2.20(s, 3H), 2.10 (s, 3H), 1.81 (m, 5H), 1.56 (m, 2H), 1.40-1.45 (m, 2H),1.16-1.29 (m, 2H).

Example 59: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-(propyl(tetrahydro-2H-pyran-4-yl)amino)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (15 g,61.5 mmol) and dihydro-2H-pyran-4(3)-one (9.2 g, 92 mmol) indichloroethane (300 mL) was added acetic acid (22 g, 369 mmol) and thereaction mixture stirred at room temperature for 15 minutes, upon whichthe reaction mixture was cooled to 0° C. and sodiumtriacetoxyborohydride (39 g, 183.96 mmol) was added. The reactionmixture was stirred overnight at room temperature. Aqueous sodiumbicarbonate was then added to the reaction mixture adjusting the pH to7-8. The organic phase was separated and the aqueous phase extractedwith ethyl acetate. The combined extracts were dried over anhydroussodium sulfate, filtered and concentrated under reduced pressure. Thecrude product was purified by column chromatography (100-200 mesh silicagel) eluting with ethyl acetate: hexane to afford methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate as anoff-white solid (14 g, 69%).

Step 2: Synthesis of methyl5-bromo-2-methyl-3-(propyl(tetrahydro-2H-pyran-4-yl)amino)benzoate

To a stirred solution of5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (1 g, 3.04mmol) and propionaldehyde (0.354 g, 6.09 mmol) in dichloroethane (10mL), acetic acid (1.12 g, 18.2 mmol) was added. The reaction mixture wasstirred at room temperature for 10 minutes. Then sodiumtriacetoxyborohydride (1.94 g, 9.14 mmol) was added at 0° C. and thereaction mixture stirred at room temperature for 2 h. The solvent wasthen removed under reduced pressure and water added to the residue. Themixture was extracted with DCM. The combined extracts were dried oversodium sulfate, filtered and concentrated under reduced pressure to givethe crude product which was purified by column chromatography to affordmethyl5-bromo-2-methyl-3-(propyl(tetrahydro-2H-pyran-4-yl)amino)benzoate (0.96g, 85.7%).

Step 3: Synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(propyl(tetrahydro-2H-pyran-4-yl)amino)benzamide

Aqueous NaOH (0.156 g, 3.8 mmol) was added to a solution of5-bromo-2-methyl-3-(propyl(tetrahydro-2H-pyran-4-yl)amino)benzoate (0.96g, 2.59 mmol) in ethanol (5 mL). The reaction mixture was stirred at 60°C. for 1 h. The ethanol was then removed under reduced pressure and theresidue acidified to pH 6 using dilute HCl and to pH 4 with citric acid.The mixture was extracted with ethyl acetate. The combined extracts weredried, filtered and concentrated giving the respective acid (0.8 g,86.67%).

The above acid (0.8 g, 2.24 mmol) was dissolved in DMSO (5 mL) and3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.683 g, 4.49 mmol) wasadded. The reaction mixture was stirred at room temperature for 15 minbefore PyBOP (1.75 g, 3.36 mmol) was added to it and stirring wascontinued for overnight. The reaction mixture was poured into ice waterand extracted with 10% MeOH/DCM. The combined extracts were dried,filtered, and concentrated to obtain the crude product which purified bysolvent washings to afford5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(propyl(tetrahydro-2H-pyran-4-yl)amino)benzamide(0.9 g, 81.8%).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-(propyl(tetrahydro-2H-pyran-4-yl)amino)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(propyl(tetrahydro-2H-pyran-4-yl)amino)benzamide(0.2 g, 0.412 mmol) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine(0.148 g, 0.488 mmol) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃(0.108 g, 1.01 mmol) was added and reaction mixture purged with argonfor 15 min. Pd (PPh₃)₄ (0.048 g, 0.042 mmol) was then added and thereaction mixture again purged with argon for 10 min. The reactionmixture was heated at 100° C. for 2 h. The reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. The combined extracts weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude product which was purified by column chromatography over silicagel to afford the title compound (0.20 g, 83.68%). LCMS: 587.40 (M+1)⁺;HPLC: 98.68% (@ 254 nm) (R_(t); 4.257; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.19 (t, 1H, J=4.8Hz), 7.56 (d, 2H, J=8 Hz), 7.38 (t, 3H, J=8 Hz), 7.19 (s, 1H), 5.85 (s,1H), 4.28 (d, 2H, J=4.4 Hz), 3.82-3.85 (m, 2H), 3.57 (m, 4H), 3.48 (s,2H), 3.23 (t, 2H, J=10.8 Hz), 2.94-3.02 (m, 3H), 2.36 (bs, 4H), 2.24 (s,3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.56-1.65 (m, 4H), 1.20-1.25 (m, 2H),0.76 (t, 3H, J=6.8 Hz).

Example 60: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (1 g, 3.04mmol) and isobutyraldehyde (1.09 g, 15.24 mmol) in methanol (15 mL),acetic acid (0.456 g, 7.6 mmol) was added. The reaction mixture wasstirred at room temperature for 8 h. Sodium cyanoborohydride (0.522 g,7.56 mmol) was then added at 0° C. and the reaction mixture stirredovernight at room temperature. The solvent was then removed underreduced pressure and crude product purified by column chromatography toafford methyl5-bromo-3-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate(0.52 g, 54.33%).

Step 2: Synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

Aqueous NaOH (0.104 g, 2.61 mmol) was added to a solution of methyl5-bromo-3-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate(0.5 g, 1.30 mmol) in ethanol (15 mL) and stirred at 60° C. for 1 h. Theethanol was then removed under reduced pressure and acidified to pH 6with dilute HCl and to pH 4 with citric acid. The mixture was extractedwith ethyl acetate. The combined extracts were dried and concentratedgiving the respective acid (0.375 g, 76.9%).

The above acid (0.350 g, 9.45 mmol) was then dissolved in DMSO (5 mL)and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.283 g, 18.9 mmol)was added. The reaction mixture was stirred at room temperature for 15min before PyBOP (0.737 g, 14.17 mmol) was added. The reaction mixturewas stirred overnight. The reaction mixture was poured into ice waterand the resulting precipitate was collected and purified by solventwashings giving5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(0.2 g, 42.01%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(isobutyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(0.14 g, 0.277 mmol) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine(0.100 g, 0.333 mmol) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃(0.108 g, 1.01 mmol) was added and solution purged with argon for 15min. Pd (PPh₃)₄ (0.032 g, 0.027 mmol) was then added and the reactionmixture again purged with argon for 10 min. The reaction mixture washeated at 100° C. for 2 h. The reaction mixture was then diluted withwater and extracted with 10% MeOH/DCM. The combined extracts were driedover Na₂SO₄ and the solvent removed under reduced pressure to afford thecrude product which was purified by preparative HPLC to afford the titlecompound as a TFA salt (0.039 g, 23.49%). LCMS: 601.30 (M+1)⁺; HPLC:99.88% (@ 254 nm) (R_(t); 5.225; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30 OC; Flow rate: 1.4 mL/min.; Gradient: 5%B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 9.83 (bs, 1H), 8.20 (t, 1H), 7.73(d, 2H, J=8 Hz), 7.56 (d, 2H, J=8 Hz), 7.43 (s, 1H), 7.21 (s, 1H), 5.86(s, 1H), 4.39 (bs, 2H), 4.28 (d, 2H, J=4.4 Hz), 3.95-3.98 (m, 2H),3.85-3.87 (m, 2H), 3.62 (t, 2H, J=11.2 Hz), 3.15-3.31 (m, 9H), 2.84 (m,1H), 2.26 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.62 (bs, 2H), 1.37-1.40(m, 2H), 0.80 (d, 6H, J=6 Hz).

Example 61: Synthesis of5-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (1 g, 3.04mmol) and cyclopropanecarbaldehyde (1.06 g, 15.24 mmol) in methanol (15mL), acetic acid (0.456 g, 7.6 mmol) was added The reaction mixture wasstirred at room temperature for 8 h. Sodium cyanoborohydride (0.488 g,7.62 mmol) was then added at 0° C. and reaction mixture stirredovernight at room temperature. The solvent was then removed underreduced pressure and the crude product purified by column chromatographyto afford methyl5-bromo-3-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate(0.275 g, 23.70%).

Step 2: Synthesis of5-bromo-3-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

Aqueous NaOH (0.056 g, 1.45 mmol) was added to a solution of methyl5-bromo-3-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate(0.275 g, 0.943 mmol) in ethanol (5 mL) and stirred at 60° C. for 1 h.The ethanol was then removed under reduced pressure and acidified to pH6 with dilute HCl and to pH 4 with citric acid. The mixture wasextracted with ethyl acetate. The combined extracts were dried andconcentrated giving the respective acid (0.25 g, 93.28%).

The above acid (0.250 g, 0.68 mmol) was dissolved in DMSO (3 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.155 g, 1.02 mmol) wasadded. The reaction mixture was stirred at room temperature for 15 minbefore PyBOP (0.708 g, 1.36 mmol) was added. The reaction mixture wasstirred overnight. The reaction mixture was poured into ice water andthe resulting precipitate collected and purified by solvent washingsgiving5-bromo-3-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.25 g, 73.31%).

Step 3: Synthesis of5-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-3-((cyclopropylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.25 g, 0.499 mmol) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine(0.181 g, 0.598 mmol) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (0.19g, 1.79 mmol) was added and solution purged with argon for 15 min. Pd(PPh₃)₄ (0.057 g, 0.049 mmol) was then added and the reaction mixtureagain purged with argon for 10 min. The reaction mixture was heated at100° C. for 2 h. The reaction mixture was diluted with water andextracted with 10% MeOH/DCM. The combined extracts were dried overNa₂SO₄ and the solvent removed under reduced pressure to afford thecrude product which was purified by preparative HPLC to afford the titlecompound as a TFA salt (0.085 g, 28.52%). LCMS: 599.35 (M+1)⁺; HPLC:99.21% (@ 254 nm) (R_(t); 4.191; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient:5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.51 (s, 1H), 9.83 (bs, 1H), 8.20 (s, 1H), 7.77(d, 2H, J=6.4 Hz), 7.53-7.58 (m, 3H), 7.28 (s, 1H), 5.87 (s, 1H), 4.39(bs, 2H), 4.29 (d, 2H, J=4.4 Hz), 3.95-3.98 (m, 2H), 3.59-3.65 (m, 2H),3.31-3.21 (m, 5H), 3.05-3.16 (m, 3H), 2.93 (m, 2H), 2.32 (m, 4H), 2.21(s, 3H), 2.10 (s, 3H), 1.65 (bs, 2H), 1.50 (m, 2H), 0.66 (bs, 1H), 0.28(d, 2H, J=7.2 Hz).

Example 62: Synthesis of5-(butyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Compound 62 was prepared with the method similar to that described inExample 61.

Analytical Data of TFA salt: LCMS: 601.35 (M+1)⁺; HPLC: 99.41% (@ 254nm) (R_(t); 4.482; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.47 (s, 1H), 9.89 (bs, 1H), 8.22 (t, 1H), 7.75 (d, 2H, J=8 Hz), 7.57(d, 2H, J=8 Hz), 7.44 (s, 1H), 7.25 (s, 1H), 5.86 (s, 1H), 4.39 (bs,2H), 4.28 (d, 2H, J=4.4 Hz), 3.95-3.98 (m, 3H), 3.83-3.86 (m, 4H),3.21-3.30 (m, 4H), 3.08-3.11 (m, 4H), 2.24 (s, 3H), 2.21 (s, 3H), 2.10(s, 3H), 1.62 (m, 4H), 1.20 (m, 4H), 0.79 (t, 3H, J=6.4 Hz).

Example 63: Synthesis of5-((cyclobutylmethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Compound 63 was prepared with the method similar to that described inExample 61.

Analytical Data: LCMS: 613.35 (M+1)⁺; HPLC: 99.25% (@ 254 nm) (R_(t);4.586; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.56 (d, 2H, J=7.6 Hz), 7.41 (s, 1H), 7.37 (d,2H, J=8 Hz), 7.20 (s, 1H), 5.85 (s, 1H), 4.45 (m, 2H), 4.28 (d, 2H, J=4Hz), 3.83-3.85 (m, 2H), 3.57 (m, 3H), 3.48 (s, 2H), 3.19-3.22 (m, 2H),3.08 (bs, 2H), 2.86 (m, 1H), 2.36 (m, 4H), 2.20 (s, 6H), 2.10 (s, 3H),1.70-1.78 (m, 4H), 1.56-1.63 (m, 6H).

Example 64: Synthesis of N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(morpholinomethyl) pyridin-3-yl) benzamide

Step 1: Synthesis of N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-2-methylbenzamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

(1 g, 2.15 mmol) and (6-formylpyridin-3-yl) boronic acid (0.539 g, 2.31mmol) in dioxane/water mixture (15 mL+3 mL), Na₂CO₃ (0.82 g, 7.74 mmol)was added and solution purged with argon for 15 min. Then Pd (PPh₃)₄(0.288 g, 0.25 mmol) was added and argon was purged again for 10 min.Reaction mass was heated at 80° C. for 2 h. On completion, reactionmixture was diluted with water and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over Na₂SO₄ and solvent removed under reducedpressure to afford crude material which was purified by columnchromatography over silica gel to afford the desired compound (0.60 g,57%).

Step 2: Synthesis of N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(morpholinomethyl) pyridin-3-yl) benzamide

To a stirred solution of N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-2-methylbenzamide (0.2 g, mmol) indichloroethane (3 mL) was added morpholine (5 equiv.) in 5 mL MeOH andacetic acid (2 equiv.) and the mixture was stirred at room temperaturefor 15 minutes. Then NaBH₃CN (1.5 equiv.) was added and the reactionstirred at room temperature for 16 hours. After completion (monitored byTLC), aqueous sodium bicarbonate was added to the reaction mixture tillpH 7-8, the organic phase was separated and the aqueous phase wasextracted with ethyl acetate. The combined organic layers were driedover anhydrous sodium sulfate, filtered and concentrated under reducedpressure. The crude compound was purified by column chromatography(100-200 mesh silica gel) eluting with ethyl acetate: hexane to affordthe title compound as an off-white solid. LCMS: 574.25 (M+1)⁺; HPLC:97.17% (@ 254 nm) (R_(t); 3.906; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient:5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.75 (s, 1H), 8.20 (t, 1H), 8.01 (d,1H, J=7.2 Hz), 7.50 (d, 1H, J=7.6 Hz), 7.46 (s, 1H), 7.27 (s, 1H), 5.85(s, 1H), 4.28 (d, 2H, J=3.6 Hz), 3.81-3.83 (m, 2H), 3.59-3.61 (m, 6H),3.22-3.30 (m, 2H), 3.08-3.10 (m, 2H), 3.03 (m, 1H), 2.43 (s, 4H), 2.25(s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.65-1.67 (m, 2H), 1.51-1.53 (m,2H), 0.83 (t, 3H, J=6.4 Hz).

Example 65: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (14 g,42.68 mmol) and acetaldehyde (3.75 g, 85.36 mmol) in dichloroethane (150mL), acetic acid (15.36 g, 256.08 mmol) was added and the reactionstirred at room temperature for 20 minutes. Sodium triacetoxyborohydride(27.01 g, 128.04 mmol) was then added at 0° C. and the reaction mixturestirred at room temperature for 2 h. The solvent was then removed underreduced pressure and water added to the residue. The mixture wasextracted with DCM. The combined extracts were dried over sodium sulfateand concentrated under reduced pressure to give the crude product waspurified by column chromatography to afford methyl5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate (14 g,93.33%).

Step 2: Synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

Aqueous NaOH (2.36 g, 59.15 mmol) was added to a solution of methyl5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate (14 g,39.43 mmol) in ethanol (100 mL) and stirred at 60° C. for 1 h. Theethanol was then removed under reduced pressure and acidified to pH 6with dilute HCl and to pH 4 with citric acid. The mixture was extractedwith ethyl acetate. The combined extracts were dried and concentratedgiving the respective acid (13.9 g, 100%).

The above acid (10 g, 29.23 mmol) was dissolved in DMSO (25 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (8.8 g, 58 mmol) andtriethylamine (5.6 g, 58.4 mmol) was added. The reaction mixture wasstirred at room temperature for 15 min before PyBOP (22 g, 43.8 mmol)was added. The reaction mixture was overnight. The reaction mixture waspoured into ice water and extracted with 10% MeOH/DCM. The combinedextracts were dried and concentrated to obtain the crude product whichpurified by solvent washings to afford5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(14 g, 73.68%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(0.2 g, 0.42 mmol) and (4-((dimethylamino)methyl)phenyl)boronic acid(0.15 g, 0.505 mmol) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (0.16g, 1.51 mmol) was added and the solution purged with argon for 15 min.Pd (PPh₃)₄ (0.048 g, 0.042 mmol) was the added and the reaction mixtureagain purged with argon for 10 min. The reaction mixture was heated at100° C. for 2 h. The reaction mixture was then diluted with water andextracted with 10% MeOH/DCM. The combined extracts were dried overNa₂SO₄ and the solvent removed under reduced pressure to afford thecrude product which was purified by column chromatography over silicagel to afford the title compound (0.120 g, 53.8%). LCMS: 531.30 (M+1)⁺;HPLC: 94.88% (@ 254 nm) (R_(t); 3.949; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.19 (t, 1H, J=4.4 Hz),7.61 (d, 2H, J=8 Hz), 7.39-7.41 (m, 3H), 7.23 (s, 1H), 5.86 (s, 1H),4.28 (d, 2H, J=4.8 Hz), 3.62-3.84 (m, 4H), 3.22-3.38 (m, 2H), 3.02-3.06(m, 3H), 2.30 (bs, 6H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H),1.64-1.67 (m, 2H), 1.51-1.53 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 66: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((4-methylpiperazin-1-yl)methyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(0.2 g, 0.42 mmol) and (4-((4-methylpiperazin-1-yl)methyl)phenyl)boronicacid (0.159 g, 0.505 mmol) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃(0.16 g, 1.51 mmol) was added and the solution purged with argon for 15min. Pd (PPh₃)₄ (0.048 g, 0.042 mmol) was then added and the reactionmixture again purged with argon for 10 min. The reaction mixture washeated at 100° C. for 2 h. The reaction mixture was then diluted withwater and extracted with 10% MeOH/DCM. The combined extracts were driedover Na₂SO₄ and the solvent removed under reduced pressure to afford thecrude product which was purified by preparative HPLC to afford the titlecompound as a TFA salt (0.110 g, 44.7%). LCMS: 586.40 (M+1)⁺; HPLC:96.03% (@ 254 nm) (R_(t); 3.803; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient:5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.23 (t, 1H), 7.69 (d, 2H, J=7.6Hz), 7.47 (t, 3H, J=8 Hz), 7.29 (s, 1H), 5.87 (s, 1H), 4.28 (d, 4H, J=4Hz), 3.93 (s, 3H), 3.83-3.86 (m, 2H), 3.43 (m, 2H), 3.16-3.27 (m, 8H),2.81 (s, 3H), 2.26 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.66 (m, 2H),1.57 (m, 2H), 0.84 (t, 3H, J=6 Hz).

Example 67: Synthesis of 4′-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptan-5-ylmethyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide (0.1 g, 28%)

Step 1: Synthesis of methyl 5-bromo-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate (14 g, 43mmol) and acetaldehyde (3.75 g, 85.4 mmol) in dichloroethane (150 mL),was added acetic acid (15.36 g, 256 mmol). After stirring at roomtemperature for 20 minutes, sodium triacetoxyborohydride (27.0 g, 128mmol) was added at 0° C. The mixture was stirred at room temperature for2 h. and quenched with aqueous sodium bicarbonate. The organic phase wasseparated and the aqueous phase was extracted with dichloromethane. Thecombined organic layers were dried over sodium sulphate and concentratedunder reduced pressure to give crude material which was purified bycolumn chromatography over silica gel to afford methyl 5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzoate (14 g, 93%).

Step 2: Synthesis of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

To a stirred solution of methyl 5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzoate (14 g, 39 mmol) inethanol (100 mL) was added aqueous NaOH (2.36 g, 59.1 mmol). Afterstirring 60° C. for 1 h, ethanol was removed under reduced pressure andacidified to pH 4 using dilute HCl followed by citric acid buffersolution. The mixture was extracted with ethyl acetate, and the combinedorganic layers were dried and concentrated to afford the correspondingacid (13.9 g).

To a stirred solution of the above acid (10 g, 29 mmol), 3-(aminomethyl)-4, 6-dimethylpyridin-2(1H)-one (8.8 g, 58 mmol) andtriethylamine (5.6 g, 58 mmol) in DMSO (25 mL) was added PYBOP (22 g, 44mmol) at 0° C. After stirring overnight at room temperature, the mixturewas poured onto ice and extracted with 10% MeOH/CH₂Cl₂. The combinedorganic layers were dried and concentrated under reduced pressure toobtain crude. Trituration of the crude material with solvent afforded5-bromo-N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzamide (14g, 73%).

Step 3: Synthesis of N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-formyl-4-methyl-[1, 1′-biphenyl]-3-carboxamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (5.0 g, 10 mmol) and (4-formylphenyl) boronicacid (2.35 g, 15.8 mmol) in dioxane/water (30 mL/10 mL) was added Na₂CO₃(4.01 g, 37.9 mmol). The solution was purged with argon for 15 min.,Pd(PPh₃)₄ (1.21 g, 1.05 mmol) and the mixture was heated at 100° C. for2 h. The mixture was allowed to cool to room temperature, diluted withwater and extracted with 10% MeOH/CH₂Cl₂. The combined organic layerswere dried over sodium sulphate and the solvent removed under reducedpressure. The resulting crude material was purified by columnchromatography over silica gel to afford N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-formyl-4-methyl-[1, 1′-biphenyl]-3-carboxamide (3.5 g, 66%).

The Following Reductive Amination Procedure was Used to SynthesizeCompounds 67 Through 105

To a stirred solution of N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-formyl-4-methyl-[1, 1′-biphenyl]-3-carboxamide (1.0 mmol) therequisite amine (3.0 mmol) in dichloroethane (10 mL), was added aceticacid (6.0 mmol). After stirring at room temperature for 20 minutes,sodium triacetoxyborohydride (0.63 g, 3.0 mmol) was added at 0° C. Themixture was stirred at room temperature for 2 h. and quenched withaqueous sodium bicarbonate. The organic phase was separated and theaqueous phase extracted with dichloromethane. The combined organiclayers were dried over sodium sulfate and concentrated under reducedpressure to give crude material which was purified by columnchromatography over silica gel or by RP-HPLC to afford the product infree base or trifluoroacetate salt form.

Analytical Data of 4′-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptan-5-ylmethyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide: LCMS: 585.25 (M+1)⁺; HPLC: 99.65% (@254 nm)(R_(t); 4.019; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.45 (s, 1H), 8.19 (t, 1H), 7.55 (d, 2H, J=7.6 Hz), 7.39-7.41 (m, 3H),7.21 (s, 1H), 5.85 (s, 1H), 4.35 (s, 2H), 4.28 (d, 2H, J=4.4 Hz), 3.93(d, 2H, J=7.2 Hz), 3.82 (d, 2H, J=9.6 Hz), 3.72 (d, 2H, J=4.4 Hz),3.44-3.53 (m, 3H), 3.22-3.27 (m, 1H), 3.01-3.09 (m, 2H), 2.73 (d, 1H,J=9.2 Hz), 2.23 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.79-1.82 (m, 1H),1.51-1.67 (m, 5H), 0.82 (t, 3H, J=6.8 Hz).

Example 68: 4′-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-ylmethyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide (0.15 g, 43%)

Analytical Data: LCMS: 585.35 (M+1)⁺; HPLC: 98.99% (@ 254 nm) (R_(t);3.95; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05%TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.:30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Holdfor 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs,1H), 8.18 (s, 1H), 7.56-7.54 (m, 2H), 7.41-7.39 (m, 3H), 7.21 (s, 1H),5.87 (s, 1H), 4.34 (s, 1H), 4.28 (d, 2H, J=4.4 Hz),3.93 (d, 1H, J=7.6Hz), 3.83-3.81 (m, 2H), 3.74-3.72 (m, 2H), 3.52 (d, 1H, J=6.8 Hz.), 3.44(s, 1H), 3.28-3.22 (m, 2H), 3.09-3.08 (m, 3H), 2.73 (d, 1H J=10 Hz),2.41 (d, 1H J=10 Hz), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.79 (m,1H), 1.67-1.51 (m, 5H). 0.83 (t, 3H J=6.8 Hz).

Example 69: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(pyrrolodin-1-ylmethyl)-[1,1′-biphenyl]-3-carboxamide (0.19 g)

Analytical Data: LCMS: 557.25 (M+1)⁺; HPLC: 97.70% (@ 254 nm) (R_(t);4.075; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.19 (t, 1H, J=4.4 Hz), 7.55 (d, 2H, J=8 Hz), 7.38 (d, 2H,J=6.4 Hz), 7.35 (s, 1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4Hz), 3.81-3.84 (m, 2H), 3.58 (s, 2H), 3.22-3.27 (m, 2H), 3.06-3.09 (m,2H), 2.99-3.04 (m, 1H), 2.43 (bs, 4H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10(s, 3H), 1.64-1.69 (m, 6H), 1.51-1.56 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 70: (S)—N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-(3-hydroxypyrrolodin-1-ylmethyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide TFA salt, (0.15 g, 44%)

Analytical Data: LCMS: 573.40 (M+1)⁺; HPLC: 97.97% (@ 254 nm) (R_(t);3.965; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47(s, 1H), 10.03-10.30 (m, 1H), 8.23 (s, 1H), 7.75 (d, 2H, J=7.2 Hz), 7.60(d, 2H, J=8 Hz), 7.52 (s, 1H), 7.32 (s, 1H), 5.87 (s, 1H), 4.45-4.46 (m,2H), 4.39-4.40 (m, 2H), 4.29 (d, 2H, J=5.2 Hz), 3.83-3.86 (m, 2H),3.43-3.55 (m, 2H), 3.01-3.36 (m, 6H), 2.32-2.37 (m, 2H), 2.27 (s, 3H),2.21 (s, 3H), 2.11 (s, 3H), 1.67 (m, 2H), 1.58 (m, 2H), 0.84 (t, 3H,J=6.4 Hz).

Example 71: (R)—N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-(3-hydroxypyrrolodin-1-ylmethyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.125 g, 55%)

Analytical Data: LCMS: 573.40 (M+1)⁺; HPLC: 96.12% (@ 254 nm) (R_(t);3.921; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.56 (d, 2H, J=7.6 Hz), 7.39 (s, 1H), 7.36 (d,2H, J=8 Hz), 7.21 (s, 1H), 5.85 (s, 1H), 4.68 (s, 1H), 4.28 (d, 2H, J=4Hz), 4.19 (bs, 1H), 3.81-3.84 (m, 2H), 3.56-3.59 (m, 2H), 3.22-3.25 (m,2H), 3.08-3.09 (m, 2H), 3.01 (m, 1H), 2.57-2.67 (m, 2H), 2.32 (m, 2H),2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.97-2.00 (m, 1H), 1.64-1.67(m, 2H), 1.51-1.53 (m, 3H), 0.83 (t, 3H, J=6.4 Hz).

Example 72: (S)—N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-(3-fluoropyrrolodin-1-ylmethyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide (0.05 g)

Analytical Data: LCMS: 575.35 (M+1)⁺; HPLC: 98.44% (@ 254 nm) (R_(t);4.081; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.56 (d, 2H, J=7.6 Hz), 7.39 (s, 1H), 7.34 (d,2H, J=8 Hz), 7.21 (s, 1H), 5.85 (s, 1H), 5.09-5.25 (m, 1H), 4.28 (d, 2H,J=4 Hz), 3.81-3.86 (m, 2H), 3.65 (s, 2H), 3.53-3.55 (m, 2H), 3.17-3.25(m, 2H), 3.07-3.16 (m, 7H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H),1.64-1.67 (m, 2H), 1.51-1.53 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 73: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(piperazin-1-ylmethyl)-[1, 1′-biphenyl]-3-carboxamideTFA salt, (0.18 g, 50%)

Analytical Data: LCMS: 572.10 (M+1)⁺; HPLC: 96.61% (@ 254 nm) (R_(t);3.736; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.81 (bs, 2H), 8.20 (s, 1H), 7.66 (d, 2H, J=7.2 Hz), 7.47 (d,2H, J=7.6 Hz), 7.42 (m, 1H), 7.25 (s, 1H), 5.86 (s, 1H), 4.28 (d, 2H,J=4 Hz), 3.82-3.85 (m, 4H), 3.11-3.27 (m, 9H), 2.88 (m, 4H), 2.25 (s,3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.65 (m, 2H), 1.53-1.55 (m, 2H), 0.83(t, 3H, J=6 Hz).

Example 74: (R)—N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-(3-fluoropyrrolodin-1-ylmethyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.07 g, 31%)

Analytical Data: LCMS: 575.35 (M+1)⁺; HPLC: 97.53% (@ 254 nm) (R_(t);4.079; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.57 (d, 2H, J=7.6 Hz), 7.38 (d, 2H, J=4.4 Hz),7.36 (s, 1H), 7.22 (s, 1H), 5.85 (s, 1H), 5.12-5.26 (m, 1H), 4.28 (d,2H, J=4 Hz), 3.81-3.84 (m, 2H), 3.63 (s, 2H), 3.22-3.25 (m, 2H),3.08-3.09 (m, 2H), 3.02 (m, 1H), 2.73-2.83 (m, 2H), 2.32 (m, 1H), 2.24(s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.89 (m, 1H), 1.64-1.67 (m, 2H),1.51-1.53 (m, 2H), 0.83 (t, 3H, J=7.2 Hz).

Example 75: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(piperidin-1-ylmethyl)-[1,1′-biphenyl]-3-carboxamide, (0.1 g, 88%)

Analytical Data: LCMS: 571.25 (M+1)⁺; HPLC: 98.25% (@ 254 nm) (R_(t);4.147; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.19 (t, 1H, J=5.2 Hz), 7.55 (d, 2H, J=8 Hz), 7.39 (s, 1H),7.34 (d, 2H, J=8 Hz), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4Hz), 3.82-3.83 (m, 2H), 3.43 (s, 2H), 3.24 (t, 2H, J=11.2 Hz), 3.06-3.09(m, 2H), 2.99-3.01 (m, 1H), 2.32 (bs, 4H), 2.24 (s, 3H), 2.20 (s, 3H),2.10 (s, 3H), 1.64-1.67 (m, 2H), 1.47-1.56 (m, 6H), 1.38-1.39 (m, 2H),0.83 (t, 3H, J=7.2 Hz).

Example 76: (S)—N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-((3-hydroxypiperidin-1-yl)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.25 g, 71.4%)

Analytical Data: LCMS: 587.40 (M+1)⁺; HPLC: 97.63% (@ 254 nm) (R_(t);3.997; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H, J-4.8 Hz), 7.56 (d, 2H, J=8 Hz), 7.39 (s, 1H),7.34 (d, 2H, J=8 Hz), 7.21 (s, 1H), 5.85 (s, 1H), 4.55 (d, 1H, J=4.8Hz), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.84 (m, 2H), 3.51-3.54 (m, 2H),3.43-3.45 (m, 1H), 3.06-3.09 (m, 3H), 2.99-3.01 (m, 2H), 2.79 (d, 1H,J=6.8 Hz), 2.65 (d, 1H, J=10.8 Hz), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s,3H), 1.78-1.88 (m, 2H), 1.58-1.71 (m, 2H), 1.39-1.51 (m, 4H), 1.04-1.10(m, 1H), 0.83 (t, 3H, J=6.8 Hz).

Example 77: (R)—N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-((3-hydroxypiperidin-1-yl)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.11 g, 48.6%)

Analytical Data: LCMS: 587.45 (M+1)⁺; HPLC: 98.65% (@ 254 nm) (R_(t);3.976; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.44(s, 1H), 8.18 (t, 1H), 7.56 (d, 2H, J=7.6 Hz), 7.39 (s, 1H), 7.35 (d,2H, J=8 Hz), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz),3.81-3.84 (m, 2H), 3.40-3.54 (m, 3H), 3.22-3.25 (m, 2H), 3.08-3.09 (m,2H), 3.02 (m, 1H), 2.78-2.80 (m, 2H), 2.66 (m, 2H), 2.24 (s, 3H), 2.20(s, 3H), 2.10 (s, 3H), 1.80-1.86 (m, 3H), 1.53-1.67 (m, 3H), 1.40-1.51(m, 3H), 1.04-1.06 (m, 1H), 0.83 (t, 3H, J=6.8 Hz).

Example 78: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-((4-hydroxypiperidin-1-yl)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.2 g, 57%)

Analytical Data: LCMS: 587.20 (M+1)⁺; HPLC: 99.89% (@ 254 nm) (R_(t);1.456; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.19 (t, 1H, J=4.4 Hz), 7.56 (d, 2H, J=8 Hz), 7.39 (s, 1H),7.34 (d, 2H, J=8 Hz), 7.21 (s, 1H), 5.85 (s, 1H), 4.53 (d, 1H, J=3.6Hz), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.84 (m, 2H), 3.44 (s, 3H), 3.22-3.27(m, 2H), 3.07-3.09 (m, 2H), 3.01-3.06 (m, 1H), 2.66 (m, 2H), 2.24 (s,3H), 2.20 (s, 3H), 2.10 (s, 3H), 2.00-2.04 (m, 2H), 1.64-1.67 (m, 4H),1.51-1.53 (m, 2H), 1.36-1.39 (m, 2H), 0.83 (t, 3H, J=7.2 Hz).

Example 79: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-((3-fluoropiperidin-1-yl)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.2 g, 56%)

Analytical Data: LCMS: 589.35 (M+1)⁺; HPLC: 96.06% (@ 254 nm) (R_(t);4.092; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.57 (d, 2H, J=7.2 Hz), 7.40 (s, 1H), 7.35 (d,2H, J=7.2 Hz), 7.22 (s, 1H), 5.85 (s, 1H), 4.56-4.68 (m, 1H), 4.28 (d,2H), 3.81-3.84 (m, 2H), 3.52 (s, 2H), 3.22-3.28 (m, 3H), 3.08-3.09 (m,2H), 3.02 (m, 1H), 2.65-2.72 (m, 1H), 2.39 (m, 2H), 2.24 (s, 3H), 2.20(s, 3H), 2.10 (s, 3H), 1.78-1.81 (m, 2H), 1.64-1.68 (m, 2H), 1.50-1.53(m, 4H), 0.83 (t, 3H).

Example 80: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-((4-fluoropiperidin-1-yl)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.09 g, 25%)

Analytical Data: LCMS: 589.30 (M+1)⁺; HPLC: 95.46% (@ 254 nm) (R_(t);4.156; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.56 (d, 2H), 7.39 (s, 1H), 7.37 (d, 2H), 7.21(s, 1H), 5.85 (s, 1H), 4.62-4.74 (m, 1H), 4.28 (d, 2H, J=3.2 Hz),3.81-3.84 (m, 2H), 3.49 (s, 2H), 3.22-3.25 (m, 3H), 3.08-3.09 (m, 3H),3.02 (m, 1H), 2.32 (m, 2H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H),1.82-1.85 (m, 2H), 1.64-1.67 (m, 4H), 1.51-1.53 (m, 2H), 0.83 (t, 3H,J=6.4 Hz).

Example 81: 4′-((4,4-difluoropiperidin-1-yl)methyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide, (0.1 g, 27%)

Analytical Data: LCMS: 607.35 (M+1)⁺; HPLC: 95.48% (@ 254 nm) (R_(t);4.237; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.19 (t, 1H), 7.58 (d, 2H, J=8 Hz), 7.38 (d, 2H, J=3.6 Hz),7.36 (s, 1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz),3.81-3.84 (m, 2H), 3.56 (s, 2H), 3.22-3.27 (m, 2H), 3.08-3.09 (m, 2H),2.99-3.01 (m, 1H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.90-1.99(m, 4H), 1.64-1.67 (m, 2H), 1.48-1.56 (m, 2H), 0.83 (t, 3H, J=6.4 Hz).[4H merged in solvent peak].

Example 82: 4′-(azetidin-1-ylmethyl)-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 543.40 (M+1)⁺; HPLC: 96.50% (@ 254 nm) (R_(t);4.010; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.19 (t, 1H, J=4.4 Hz), 7.54 (d, 2H, J=8 Hz), 7.38 (s, 1H),7.32 (d, 2H, J=8 Hz), 7.20 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4Hz), 3.81-3.84 (m, 2H), 3.52 (s, 2H), 3.22-3.27 (m, 2H), 2.98-3.11 (m,7H), 2.23 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.94-2.01 (m, 2H),1.64-1.67 (m, 2H), 1.51-1.56 (m, 2H), 0.82 (t, 3H, J=7.2 Hz).

Example 83: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-((3-hydroxyazetidin-1-yl)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 559.80 (M+1)⁺; HPLC: 96.10% (@ 254 nm) (R_(t);3.917; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.54 (d, 2H, J=8 Hz), 7.38 (s, 1H), 7.31 (d, 2H,J=4.4 Hz), 7.20 (s, 1H), 5.85 (s, 1H), 5.28 (d, 1H, J=6.4 Hz), 4.28 (d,2H, J=4.4 Hz), 4.17-4.19 (m, 1H), 3.81-3.84 (m, 2H), 3.56 (s, 2H), 3.48(t, 2H, J=6.4 Hz), 3.22-3.27 (m, 2H), 3.06-3.09 (m, 2H), 3.01 (m, 1H),2.75 (t, 2H, J=6.8 Hz), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H),1.64-1.67 (m, 2H), 1.51-1.53 (m, 2H), 0.82 (t, 3H, J=7.2 Hz).

Example 84: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4′-((3-fluoroazetidin-1-yl)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 561.25 (M+1)⁺; HPLC: 97.99% (@ 254 nm) (R_(t);4.021; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H), 7.57 (d, 2H, J=7.2 Hz), 7.37-7.40 (m, 3H), 7.22(s, 1H), 5.85 (s, 1H), 5.27 (m, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.84(m, 2H), 3.63 (s, 2H), 3.22-3.27 (m, 2H), 3.08-3.09 (m, 2H), 3.01 (m,1H), 2.77 (m, 2H), 2.24 (s, 3H), 2.20 (s, 3H), 2.15 (s, 3H), 1.65-1.67(m, 2H), 1.51-1.53 (m, 2H), 1.04-1.06 (m, 1H), 0.83 (t, 3H, J=7.2 Hz)[2H merged in solvent peak].

Example 86: 4′-((1,4-diazepan-1-yl)methyl)-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 585.37 (M+1)⁺; HPLC: 87.74% (@ 254 nm) (R_(t);3.715; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47(s, 1H), 8.18 (t, 1H), 7.57 (d, 2H, J=6.8 Hz), 7.39 (m, 3H), 7.21 (s,1H), 5.85 (s, 1H), 4.29 (d, 2H), 3.81-3.84 (m, 2H), 3.66 (s, 2H),3.08-3.09 (m, 3H), 3.02 (bs, 4H), 2.96 (m, 3H), 2.64-2.66 (m, 4H), 2.24(s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.78 (m, 2H), 1.64-1.67 (m, 2H),1.51-1.53 (m, 2H), 0.83 (t, 3H).

Example 87: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((4-methyl-1,4-diazepan-1-yl)methyl)-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 600.30 (M+1)⁺; HPLC: 99.46% (@ 254 nm) (R_(t);3.713; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.44(s, 1H), 8.17 (t, 1H), 7.56 (d, 2H, J=8 Hz), 7.38 (d, 2H, J=5.6 Hz),7.36 (s, 1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=5.2 Hz),3.81-3.84 (m, 2H), 3.61 (s, 2H), 3.09-3.28 (m, 3H), 3.06-3.09 (m, 2H),3.02 (m, 1H), 2.59-2.65 (m, 5H), 2.56 (t, 2H, J=6 Hz), 2.24 (s, 6H),2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.72 (m, 4H), 1.51-1.53 (m, 2H), 0.83(t, 3H, J=6.4 Hz).

Example 88: 4′-((1,4-oxazepan-4-yl)methyl)-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 587.40 (M+1)⁺; HPLC: 96.85% (@ 254 nm) (R_(t);4.055; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H, J=4.8 Hz), 7.56 (d, 2H, J=8 Hz), 7.37-7.39 (m,3H), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.84 (m,2H), 3.69 (t, 3H, J=6 Hz), 3.64 (s, 1H), 3.59-3.61 (m, 2H), 3.22-3.27(m, 2H), 2.99-3.09 (m, 3H), 2.59-2.64 (m, 4H), 2.24 (s, 3H), 2.20 (s,3H), 2.10 (s, 3H), 1.77-1.83 (m, 2H), 1.64-1.67 (m, 2H), 1.48-1.56 (m,2H), 0.83 (t, 3H, J=7.2 Hz).

Example 89: 4′-(aminomethyl)-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 503.40 (M+1)⁺; HPLC: 79.83% (@ 254 nm) (R_(t);3.846; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.19 (t, 1H), 7.63 (d, 2H, J=8.4 Hz), 7.47 (d, 2H, J=8 Hz),7.39 (s, 1H), 7.23 (s, 1H), 5.86 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.92(s, 2H), 3.81-3.84 (m, 2H), 3.22-3.32 (m, 2H), 3.08-3.10 (m, 2H), 3.01(m, 1H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.65-1.67 (m, 2H),1.51-1.56 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 90: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((methylamino)methyl)-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 517.30 (M+1)⁺; HPLC: 98.05% (@ 254 nm) (R_(t);3.886; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.51(s, 1H), 8.19 (t, 1H, J=4.4 Hz), 7.55 (d, 2H, J=8 Hz), 7.38 (d, 2H),7.36 (s, 1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz),3.81-3.84 (m, 2H), 3.66 (s, 2H), 3.11-3.25 (m, 3H), 3.04-3.09 (m, 2H),2.99-3.01 (m, 1H), 2.26 (s, 3H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s,3H), 1.65-1.67 (m, 2H), 1.48-1.56 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 91: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((ethylamino)methyl)-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 531.35 (M+1)⁺; HPLC: 98.28% (@ 254 nm) (R_(t);3.977; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H, J=4.8 Hz), 7.55 (d, 2H, J=8 Hz), 7.39 (s, 1H),7.37 (d, 2H, J=2 Hz), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=6 Hz),3.81-3.84 (m, 2H), 3.71 (s, 2H), 3.22-3.28 (m, 2H), 3.01-3.11 (m, 3H),2.52-2.55 (m, 2H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.65-1.67(m, 2H), 1.50-1.53 (m, 2H), 1.03 (t, 3H, J=7.2 Hz), 0.83 (t, 3H, J=6.8Hz).

Example 92: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((isopropylamino)methyl)-[1,1′-biphenyl]-3-carboxamide TFA salt

Analytical Data of TFA salt: LCMS: 545.40 (M+1)⁺; HPLC: 94.74% (@ 254nm) (R_(t); 4.081; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.47 (s, 1H), 8.66 (bs, 1H), 8.20 (s, 1H), 7.74 (d, 2H, J=7.6 Hz),7.57 (d, 2H, J=7.6 Hz), 7.43 (s, 1H), 7.27 (s, 1H), 5.86 (s, 1H), 4.28(d, 2H, J=4.4 Hz), 4.19 (t, 2H), 3.82-3.85 (m, 2H), 3.25 (t, 2H, J=10.8Hz), 3.09-3.22 (m, 3H), 2.25 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.65(m, 2H), 1.53-1.55 (m, 3H), 1.28 (d, 6H, J=6.4 Hz), 0.83 (t, 3H, J=6.8Hz).

Example 93: 4′-((cyclopropylmethyl)amino)methyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 557.35 (M+1)⁺; HPLC: 96.44% (@ 254 nm) (R_(t);4.182; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.19 (t, 1H), 7.55 (d, 2H, J=7.2 Hz), 7.37-7.39 (m, 3H), 7.21(s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.84 (m, 2H), 3.73(s, 2H), 3.22-3.24 (m, 3H), 3.06-3.09 (m, 2H), 3.01 (m, 1H), 2.36 (d,2H, J=6.8 Hz), 2.23 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m,2H), 1.51-1.56 (m, 2H), 0.81-0.084 (m, 4H), 0.38-0.39 (m, 2H), 0.07-0.08(m, 2H).

Example 94: 4′-((diethyl)amino)methyl)-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 559.20 (M+1)⁺; HPLC: 98.33% (@ 254 nm) (R_(t);4.126; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47(s, 1H), 8.19 (t, 1H, J=5.2 Hz), 7.55 (d, 2H, J=8 Hz), 7.38 (d, 2H,J=5.2 Hz), 7.36 (s, 1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4Hz), 3.81-3.83 (m, 2H), 3.54 (s, 2H), 3.22-3.37 (m, 2H), 3.06-3.11 (m,2H), 2.99-3.01 (m, 1H), 2.43-2.47 (m, 4H), 2.24 (s, 3H), 2.20 (s, 3H),2.10 (s, 3H), 1.64-1.67 (m, 2H), 1.48-1.56 (m, 2H), 0.98 (t, 6H, J=7.2Hz), 0.83 (t, 3H, J=6.8 Hz).

Example 95: (R)-4′-(((2,3-dihydroxypropyl)amino)methyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 599.35 (M+1)⁺; HPLC: 93.58% (@ 254 nm) (R_(t);3.808; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H, J=4.8 Hz), 7.56 (d, 2H, J=8 Hz), 7.37-7.39 (m,3H), 7.16 (s, 1H), 5.85 (s, 1H), 4.51-4.56 (m, 2H), 4.28 (d, 2H, J=4.8Hz), 3.81-3.84 (m, 2H), 3.73 (s, 2H), 3.55 (m, 1H), 3.11-3.25 (m, 3H),3.01-3.09 (m, 3H), 2.56-2.61 (m, 1H), 2.41-2.46 (m, 2H), 2.24 (s, 3H),2.20 (s, 3H), 2.15 (s, 3H), 1.65-1.67 (m, 2H), 1.48-1.56 (m, 2H), 0.83(t, 3H, J=6.8 Hz).

Example 96: (S)-4′-(((2,3-dihydroxypropyl)amino)methyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 577.25 (M+1)⁺; HPLC: 96.96% (@ 254 nm) (R_(t);3.812; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.19 (t, 1H, J=4.8 Hz), 7.55 (d, 2H, J=8 Hz), 7.37-7.39 (m,3H), 7.21 (s, 1H), 5.85 (s, 1H), 4.55 (m, 3H), 4.28 (d, 2H, J=4.8 Hz),3.81-3.83 (m, 2H), 3.72 (s, 2H), 3.55 (bs, 1H), 3.22-3.28 (m, 3H),3.01-3.11 (m, 3H), 2.57-2.60 (m, 1H), 2.41-2.45 (m, 2H), 2.24 (s, 3H),2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m, 2H), 1.51-1.56 (m, 2H), 0.83(t, 3H, J=7.2 Hz).

Example 97:4′-(((cyclopropylmethyl)(methyl)amino)methyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 571.40 (M+1)⁺; HPLC: 99.80% (@ 254 nm) (R_(t);4.243; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(s, 1H), 8.18 (t, 1H, J=4.8 Hz), 7.57 (d, 2H, J=8 Hz), 7.40 (s, 1H),7.37 (d, 2H, J=8 Hz), 7.22 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4Hz), 3.81-3.83 (m, 2H), 3.53 (s, 2H), 3.11-3.25 (m, 2H), 2.99-3.09 (m,3H), 2.25-2.32 (m, 2H), 2.24 (s, 3H), 2.20 (s, 6H), 2.10 (s, 3H),1.65-1.67 (m, 2H), 1.48-1.56 (m, 2H), 0.88 (m, 1H), 0.83 (t, 3H, J=7.2Hz), 0.46-0.47 (m, 2H), 0.081 (m, 2H).

Example 98: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(((2-hydroxyethyl)amino)methyl)-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 547.35 (M+1)⁺; HPLC: 96.46% (@ 254 nm) (R_(t);3.862; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.44(s, 1H), 8.19 (t, 1H, J=4.4 Hz), 7.55 (d, 2H, J=8 Hz), 7.38 (d, 2H,J=4.4 Hz), 7.37 (s, 1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.47 (bs, 2H), 4.28(d, 2H, J=3.6 Hz), 3.81-3.83 (m, 2H), 3.72 (s, 2H), 3.46 (m, 2H),3.22-3.27 (m, 2H), 3.07-3.09 (m, 2H), 3.01-3.06 (m, 1H), 2.55-2.57 (m,2H), 2.23 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m, 2H),1.51-1.53 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 99: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(((3-hydroxypropyl)amino)methyl)-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 561.30 (M+1)⁺; HPLC: 96.82% (@ 254 nm) (R_(t);3.911; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47(s, 1H), 8.19 (t, 1H), 7.55 (d, 2H, J=7.6 Hz), 7.39 (d, 2H), 7.37 (s,1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.46 (bs, 1H), 4.28 (d, 2H, J=4.4 Hz),3.81-3.83 (m, 2H), 3.69 (s, 2H), 3.45 (t, 2H, J=6.4 Hz), 3.22-3.27 (m,2H), 3.07-3.09 (m, 2H), 3.01-3.06 (m, 1H), 2.23 (s, 3H), 2.20 (s, 3H),2.14 (s, 3H), 1.64-1.67 (m, 2H), 1.51-1.61 (m, 4H), 0.83 (t, 3H, J=6.8Hz) [1H merged in solvent peak].

Example 100: 4′-((bis(2-hydroxyethyl)amino)methyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide TFA salt

Analytical Data TFA salt: LCMS: 591.25 (M+1)⁺; HPLC: 99.00% (@ 254 nm)(R_(t); 3.860; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.47 (s, 1H), 9.38 (s, 1H), 8.25 (s, 1H), 7.77 (d, 2H, J=3.2 Hz), 7.65(s, 2H), 7.63 (s, 1H), 7.33 (s, 1H), 5.87 (s, 1H), 4.46 (s, 2H), 4.29(d, 2H, J=4.8 Hz), 3.78-3.90 (m, 6H), 3.18-3.28 (m, 9H), 2.27 (s, 3H),2.21 (s, 3H), 2.10 (s, 3H), 1.58-1.67 (m, 4H), 0.85 (t, 3H, J=6.8 Hz).

Example 101: 4′-(((2-aminoethyl)amino)methyl)-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1, 1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 546.35 (M+1)⁺; HPLC: 93.12% (@ 254 nm) (R_(t);3.721; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47(s, 1H), 9.19 (bs, 1H), 8.20 (t, 1H), 7.99 (bs, 2H), 7.74 (d, 2H, J=8Hz), 7.57 (d, 2H, J=8 Hz), 7.42 (s, 1H), 7.26 (s, 1H), 5.86 (s, 1H),4.26-4.29 (m, 3H), 3.82-3.84 (m, 2H), 3.11-3.27 (m, 8H), 3.03 (s, 2H),2.24 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m, 2H), 1.55 (m,2H), 0.83 (t, 3H, J=6 Hz).

Example 102: 4′-(((3-aminopropyl)amino)methyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide TFA salt

Analytical Data TFA salt: LCMS: 560.20 (M+1)⁺; HPLC: 98.90% (@ 254 nm)(R_(t); 3.611; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.48 (s, 1H), 8.93 (bs, 2H), 8.20 (t, 1H), 7.79 (bs, 2H), 7.73 (d, 2H,J=8 Hz), 7.55 (d, 2H, J=8 Hz), 7.43 (s, 1H), 7.27 (s, 1H), 5.87 (s, 1H),4.28 (d, 2H, J=4.4 Hz), 4.19 (m, 2H), 3.81-3.85 (m, 2H), 3.25 (t, 2H,J=11.2 Hz), 3.11-3.16 (m, 3H), 3.01 (m, 3H), 2.87-2.88 (m, 1H), 2.24 (s,3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.89-1.92 (m, 2H), 1.65-1.68 (m, 2H),1.53-1.55 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 103: 4′-(((2,2-difluoroethyl)amino)methyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 567.30 (M+1)⁺; HPLC: 92.86% (@ 254 nm) (R_(t);3.984; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.19 (t, 1H), 7.57 (d, 2H, J=7.2 Hz), 7.38-7.40 (m, 3H), 7.21(s, 1H), 6.01 (t, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.84(m, 2H), 3.77 (s, 2H), 3.22-3.28 (m, 2H), 3.06-3.09 (m, 2H), 3.01 (m,1H), 2.84 (t, 2H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.65-1.67(m, 2H), 1.51-1.53 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 104: N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(((2,2,2-trifluoroethyl)amino)methyl)-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 585.25 (M+1)⁺; HPLC: 99.52% (@ 254 nm) (R_(t);4.175; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 8.19 (t, 1H), 7.58 (d, 2H, J=8 Hz), 7.39-7.40 (m, 3H), 7.21 (s,1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.80-3.83 (m, 4H), 2.93-3.27(m, 8H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m, 2H),1.51-1.53 (m, 2H), 0.83 (t, 3H, J=6.8 Hz).

Example 105: 4′-(2-oxa-6-azaspiro[3.3]heptan-6-ylmethyl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 585.40 (M+1)⁺; HPLC: 99.67% (@ 254 nm) (R_(t);3.99; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05%TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.:30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Holdfor 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs,1H), 8.18 (s, 1H), 7.55-7.54 (m, 2H), 7.38 (s, 1H), 7.31-7.29 (m, 2H),7.20 (s, 1H), 5.85 (s, 1H), 4.60 (s, 3H), 4.28 (d, 2H, J=4.8 Hz),3.83-3.81 (m, 2H), 3.53 (s, 2H), 3.83-3.81 (m, 2H), 3.32 (2 Protonsmerged in solvent peak), 3.24-3.22 (m, 4H), 3.09-3.01 (m, 3H), 2.24 (s,3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.67-1.64 (m, 2H), 1.53-1.51 (m, 2H),0.83 (t, 3H J=6.4 Hz).

Example 108: Synthesis of5-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(methyl)amino)-cyclohexyl)carbamate(1 equiv.) andN,N-dimethyl-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanamine(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and the reaction flask was purged again for 10 min.with argon. The reaction mixture was heated at 100° C. for 4 h. Thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by column chromatography over silica gel to afford tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate(0.08 g, 48.78%)

Step 2: Synthesis of5-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

A stirred solution of tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4′-((dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(methyl)amino)cyclohexyl)carbamate(0.08 g) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. The reactionwas concentrated to dryness yielding the title compound as a TFA salt(0.06 g, 89.55%). LCMS: 530.35 (M+1)⁺; HPLC: 89.74% (@ 254 nm) (R_(t);3.557; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46(s, 1H), 9.74 (bs, 1H), 8.17 (t, 1H), 7.74-7.76 (m, 4H), 7.55 (d, 2H,J=7.6 Hz), 7.36 (s, 1H), 7.22 (s, 1H), 5.86 (s, 1H), 4.28-4.31 (m, 4H),2.97 (bs, 1H), 2.74 (d, 6H, J=4.4 Hz), 2.66 (s, 3H), 2.20 (d, 6H, J=2Hz), 2.10 (s, 3H), 1.92-1.95 (m, 2H), 1.74-1.77 (m, 2H), 1.52-1.57 (m,2H), 1.28-1.30 (m, 2H) [1H merged in solvent peak].

Example 109: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(0.2 g, 0.42 mmol) and (1-methyl-1H-pyrazol-4-yl)boronic acid (0.105 g,0.505 mmol) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (0.16 g, 1.51mmol) was added and solution purged with argon for 15 min. Pd (PPh₃)₄(0.048 g, 0.042 mmol) was then added and the reaction mixture againpurged with argon for 10 min. The reaction mixture was heated at 100° C.for 2 h. The reaction mixture was then diluted with water and extractedwith 10% MeOH/DCM. The combined extracts were dried over Na₂SO₄ and thesolvent removed under reduced pressure to afford the crude product whichwas purified by column chromatography to afford the title compound(0.100 g, 50%). LCMS: 478.20 (M+1)⁺; HPLC: 95.82% (@ 254 nm) (R_(t);4.322; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 11.45(s, 1H), 8.12 (s, 1H), 8.10 (t, 1H), 7.81 (s, 1H), 7.33 (s, 1H), 7.13(s, 1H), 5.86 (s, 1H), 4.27 (d, 2H, J=4.8 Hz), 3.81-3.83 (m, 5H),3.21-3.26 (m, 2H), 2.98-3.08 (m, 3H), 2.20 (s, 3H), 2.17 (s, 3H), 2.10(s, 3H), 1.63-1.66 (m, 2H), 1.48-1.52 (m, 2H), 0.86 (t, 3H, J=7.2 Hz).

Example 110: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-2-methylbenzamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(1 g, 2.15 mmol) and (6-formylpyridin-3-yl) boronic acid (0.539 g, 2.31mmol) in dioxane/water mixture (15 mL+3 mL), Na₂CO₃ (0.82 g, 7.74 mmol)was added and solution purged with argon for 15 min. Then Pd (PPh₃)₄(0.288 g, 0.25 mmol) was added and argon was purged again for 10 min.Reaction mass was heated at 80° C. for 2 h. On completion, reactionmixture was diluted with water and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over Na₂SO₄ and solvent removed under reducedpressure to afford crude material which was purified by columnchromatography over silica gel to afford the desired compound (0.60 g,57%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-((dimethylamino)methyl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-2-methylbenzamide(0.102 g, 0.203 mmol) and dimethylamine (0.044 g, 2M 0.507 mL, 1.01mmol) in dichloroethane (3 mL), acetic acid (0.073 g, 1.021 mmol) wasadded and reaction stirred at room temperature for 20 minutes. Thensodium triacetoxyborohydride (0.129 g, 0.609 mmol) was added at 0° C.and reaction stirred for 4 h at room temperature. On completion, solventwas removed under reduced pressure and water was added, extraction wascarried out using 10% MeOH/DCM. Combined organic layers were dried oversodium sulphate and concentrated under reduced pressure giving crudematerial which was purified by column chromatography to afford the titlecompound (0.08 g, 75%). LCMS: 532.30 (M+1)⁺; HPLC: 97.53% (@ 254 nm)(R_(t); 3.878; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (s, 1H), 8.75 (d, 1H, J=1.2 Hz), 8.20 (t, 1H, J=4.8 Hz), 8.02 (d,1H, J=6.4 Hz), 7.49 (s, 1H), 7.47 (s, 1H), 7.27 (s, 1H), 5.87 (s, 1H),4.28 (d, 2H, J=4.8 Hz), 3.81-3.84 (m, 2H), 3.56 (s, 2H), 3.22-3.24 (m,2H), 3.02-3.12 (m, 3H), 2.25 (s, 3H), 2.21 (s, 6H), 2.20 (s, 3H), 2.10(s, 3H), 1.65-1.67 (m, 2H), 1.50-1.56 (m, 2H), 0.82 (t, 3H, J=6.8 Hz).

Example 111: Synthesis of N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-((4-methylpiperazin-1-yl) methyl) pyridin-3-yl)benzamide

Step 1a: Synthesis of N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-2-methylbenzamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

(1 g, 2.15 mmol) and (6-formylpyridin-3-yl) boronic acid (0.539 g, 2.31mmol) in dioxane/water mixture (15 mL+3 mL), Na₂CO₃ (0.82 g, 7.74 mmol)was added and solution purged with argon for 15 min. Then Pd (PPh₃)₄(0.288 g, 0.25 mmol) was added and argon was purged again for 10 min.Reaction mass was heated at 80° C. for 2 h. On completion, reactionmixture was diluted with water and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over Na₂SO₄ and solvent removed under reducedpressure to afford crude material which was purified by columnchromatography over silica gel to afford the desired compound (0.60 g,57%).

Step 1b:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-(hydroxymethyl)pyridin-3-yl)-2-methylbenzamide

When the above reaction was repeated on a 1.5 g scale,N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-(hydroxymethyl)pyridin-3-yl)-2-methylbenzamidewas isolated (0.350 g, 22%).

Step 2: Synthesis of 5-(6-(bromomethyl)pyridin-3-yl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-(hydroxymethyl)pyridin-3-yl)-2-methylbenzamide(0.35 g, 0.694 mmol) in DCM (5 mL), triphenyl phosphine (0.361 g, 1.38mmol) was added and stirred it at room temperature for 10 min. FinallyCBr₄ (0.318 g, 1.38 mmol) was added portion wise to it and resultingsolution was stirred at room temperature for 18 h. On completion, waterwas added to the reaction mass and extraction was carried out using DCM.Combined organic layers were dried over sodium sulphate, concentratedunder reduced pressure to give crude material which then columnpurification gave desired compound (0.35 g, 89%).

Step 3: Synthesis of N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-((4-methylpiperazin-1-yl) methyl) pyridin-3-yl)benzamide

To stirred solution of 5-(6-(bromomethyl) pyridin-3-yl)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzamide (0.175 g, 0.309mmol) dissolved in THF (2 mL), was added 1-methyl-piperazine (0.309 g,1.54 mmol) at room temperature and stirred at the same temperature for18 h. On completion, water was added to the reaction mass and extractionwas carried out using DCM. Combined organic layers were dried oversodium sulphate and concentrated under reduced pressure giving crudematerial which then purified by preparative HPLC to give the titlecompound as a TFA salt (0.028 g, 15%). LCMS: 587.40 (M+1)⁺; HPLC: 98.05%(@ 254 nm) (R_(t); 3.831; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% Bto 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.46 (s, 1H), 8.89 (s, 1H), 8.21 (d, 2H, J=7.6 Hz), 7.59 (d,2H, J=7.6 Hz), 7.35 (s, 1H), 5.87 (s, 1H), 4.29 (d, 2H, J=4 Hz),3.96-4.04 (m, 2H), 3.83-3.86 (m, 2H), 3.16-3.43 (m, 13H), 2.81 (s, 3H),2.27 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.67 (m, 2H), 1.56 (m, 2H),0.84 (t, 3H, J=6.4 Hz).

Example 112: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4,4′-dimethyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (200 mg, 0.42 mmol) and p-tolyl boronic acid(86 mg, 0.63 mmol) in dioxane (3 mL), aqueous 2M Na₂CO₃ solution (0.75mL, 1.51 mmol) was added and solution was purged with argon for 15 min.Then Pd(PPh₃)₄ (48 mg, 0.04 mmol) was added and argon was purged againfor 15 min. Reaction mass was heated at 100° C. for 2 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the title compound (150 mg, 73%).LCMS: 488.20 (M+1)⁺; HPLC: 99.33% (@ 254 nm) (R_(t); 5.393; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.19(t, 1H), 7.51 (d, 2H, J=8 Hz), 7.37 (s, 1H), 7.25 (d, 2H, J=8 Hz), 7.19(s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.83 (m, 2H),3.22-3.27 (m, 2H), 3.07-3.09 (m, 2H), 3.01 (m, 1H), 2.33 (s, 3H), 2.23(s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m, 2H), 1.51-1.55 (m,2H), 0.82 (t, 3H, J=6.8 Hz).

Example 113: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4′-(hydroxymethyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(200 mg, 0.42 mmol) and 4-(hydroxymethyl)phenylboronic acid (96 mg, 0.63mmol) in dioxane (2.5 mL), aqueous 2M Na₂CO₃ solution (0.75 mL, 1.51mmol) was added and solution was purged with argon for 15 min. ThenPd(PPh₃)₄ (48 mg, 0.04 mmol) was added and argon was purged again for 15min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the title compound (130 mg, 62%).LCMS: 504.15 (M+1)⁺; HPLC: 98.86% (@ 254 nm) (R_(t); 4.240; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (s, 1H), 8.19(t, 1H), 7.57 (d, 2H, J=7.2 Hz), 7.39 (s, 1H), 7.37 (d, 2H), 7.21 (s,1H), 5.85 (s, 1H), 5.20 (t, 1H, J=5.2 Hz), 4.52 (d, 2H, J=5.6 Hz), 4.28(d, 2H, J=3.6 Hz), 3.81-3.84 (m, 2H), 3.22-3.32 (m, 2H), 3.08-3.09 (m,2H), 3.01 (m, 1H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.65-1.67(m, 2H), 1.51-1.53 (m, 2H), 0.83 (t, 3H, J=6.4 Hz).

Example 114: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-3′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-3′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(400 mg, 0.84 mmol) and (3-formylphenyl)boronic acid (189 mg, 1.26 mmol)in dioxane (2 mL), aqueous 2M Na₂CO₃ solution (1.5 mL, 3.03 mmol) wasadded and solution was purged with argon for 15 min. Then Pd(PPh₃)₄ (97mg, 0.08 mmol) was added and argon was purged again for 15 min. Reactionmass was heated at 100° C. for 4 h. On completion, reaction mixture wasdiluted with water and extracted with 10% MeOH/DCM (3 times). Combinedorganic layer was dried over sodium sulphate. Removal of the solventunder reduced pressure followed by column chromatographic purificationafforded the title compound (270 mg, 64%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-3′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-3′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide(270 mg, 0.53 mmol) and morpholine (94 mg, 1.07 mmol) in dichloroethane(5 mL), acetic acid (194 mg, 3.23 mmol) was added and reaction stirredat room temperature for 30 minutes. Then sodium triacetoxyborohydride(343 mg, 1.61 mmol) was added to the reaction mixture at 0° C., allowedto attain room temperature and stirring continued for overnight. Oncompletion, reaction mixture was diluted with dichloromethane, washedwith water, saturated aqueous sodium bicarbonate solution and dried oversodium sulphate. Removal of the solvent under reduced pressure followedby column chromatographic purification afforded the title compound (200mg, 65%). LCMS: 573.25 (M+1)⁺; HPLC: 90.21% (@ 254 nm) (R_(t); 4.048;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30°C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 11.45 (s, 1H),8.20 (t, 1H), 7.52 (s, 1H), 7.49 (d, 2H, J=7.6 Hz), 7.39 (d, 1H, J=5.6Hz), 7.29 (d, 1H, J=7.2 Hz), 7.20 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H,J=4.4 Hz), 3.82-3.84 (m, 2H), 3.56 (m, 4H), 3.52 (s, 2H), 3.22-3.30 (m,2H), 3.08-3.10 (m, 2H), 3.01 (m, 1H), 2.37 (s, 4H), 2.24 (s, 3H), 2.20(s, 3H), 2.10 (s, 3H), 1.65-1.67 (m, 2H), 1.51-1.54 (m, 2H), 0.83 (t,3H, J=6.4 Hz).

Example 115:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((3-(morpholinomethyl)azetidin-1-yl)methyl)-[1,1′-biphenyl]-3-carboxamide

Compound 115 was prepared with the method similar to that described inExample 67. Analytical Data: LCMS: 642.45 (M+1)⁺; HPLC: 93.13% (@ 254nm) (R_(t); 3.803; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.45 (s, 1H), 8.18 (t, 1H), 7.54 (d, 2H, J=7.6 Hz), 7.38 (s, 1H),7.31 (d, 2H, J=7.6 Hz), 7.20 (s, 1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4Hz), 3.81-3.83 (m, 2H), 3.52-3.53 (m, 6H), 3.22-3.24 (m, 2H), 3.07-3.09(m, 2H), 3.01 (m, 1H), 2.79 (s, 2H), 2.56-2.58 (m, 2H), 2.29 (m, 2H),2.28 (s, 3H), 2.23 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.64-1.67 (m,2H), 1.51-1.53 (m, 2H), 0.82 (t, 3H, J=6.8 Hz).

Example 116: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-((tetrahydro-2H-pyran-4-yl)amino)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl4-methyl-4′-(morpholinomethyl)-5-((tetrahydro-2H-pyran-4-yl)amino)-[1,1′-biphenyl]-3-carboxylate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (600 mg,1.83 mmol) and4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (833mg, 2.75 mmol) in dioxane (9 mL), aqueous 2M Na₂CO₃ solution (3.30 mL,6.60 mmol) was added and solution was purged with argon for 15 min. ThenPd(PPh₃)₄ (211 mg, 0.18 mmol) was added and argon was purged again for15 min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the title compound (500 mg, 77%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-((tetrahydro-2H-pyran-4-yl)amino)-[1,1′-biphenyl]-3-carboxamide

Aqueous NaOH (73 m g, 1.76 mmol) was added to a stirred solution ofmethyl4-methyl-4′-(morpholinomethyl)-5-((tetrahydro-2H-pyran-4-yl)amino)-[1,1′-biphenyl]-3-carboxylate(500 mg, 1.17 mmol) in ethanol (10 mL) and stirring continued at 60° C.for 1 h. After completion, ethanol was removed under reduced pressureand acidified using dilute HCl up to pH 6. Aqueous layer was extractedethyl acetate (5 times) and the combined organic layer was dried oversodium sulphate. Removal of the solvent under reduced pressure affordedrespective acid (350 mg, 72.4%).

To an stirred ice cooled solution of above acid (200 mg, 0.48 mmol) inDMF (10 mL) EDCI (139 mg, 0.73 mmol) and triethylamine (0.17 mL, 1.21mmol) were added. Then after 15 minutes interval of stirring at 0° C.,HOBT (78 mg, 0.58 mmol) followed by 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (148 mg, 0.97 mmol) were added. Reactionmixture was allowed to attain room temperature and stirring continuedfor overnight. On completion, the reaction mass was poured into ice,extracted with 10% MeOH/DCM (5 times). Combined organic layer was washedwith water and dried over sodium sulphate. Removal of the solvent underreduced pressure followed by solvent washings afforded the titlecompound (50 mg, 19%). LCMS: 545.15 (M+1)⁺; HPLC: 95.86% (@ 254 nm)(R_(t); 4.382; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.45 (s, 1H), 8.03 (m, 1H), 7.71 (bs, 1H), 7.54 (d, 1H, J=7.6 Hz), 7.34(d, 2H, J=7.6 Hz), 6.85 (s, 1H), 6.70 (s, 1H), 5.83 (d, 2H, J=7.6 Hz),4.58 (d, 1H, J=7.6 Hz), 4.26 (d, 2H, J=4 Hz), 4.04 (d, 2H, J=4.8 Hz),3.85-3.88 (m, 2H), 3.62 (m, 1H), 3.57 (t, 2H), 3.41-3.47 (m, 3H),2.32-2.36 (m, 4H), 2.19 (s, 3H), 2.11 (s, 3H), 2.05 (s, 3H), 1.88-1.91(m, 2H), 1.50-1.52 (m, 2H).

Example 117: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethylamino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl 5-bromo-3-(ethylamino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (1.0 g,4.09 mmol) and acetaldehyde (180 mg, 4.09 mmol) in dichloroethane (10mL), acetic acid (1.47 g, 24.58 mmol) was added and reaction stirred atroom temperature for 30 minutes. Then sodium triacetoxyborohydride (2.6g, 12.29 mmol) was added at 0° C., allowed to attain room temperatureand stirring continued for 2 h. On completion, reaction mixture wasdiluted with dichloromethane, washed with water, saturated aqueoussodium bicarbonate solution and dried over sodium sulphate. Removal ofthe solvent under reduced pressure followed by column chromatographicpurification afforded the desired compound (600 mg, 55%).

Step 2: Synthesis of methyl5-(ethylamino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxylate

To a stirred solution of methyl 5-bromo-3-(ethylamino)-2-methylbenzoate(600 mg, 2.2 mmol) and4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (1.0g, 3.3 mmol) in dioxane (5 mL), aqueous 2M Na₂CO₃ solution (3.96 mL,7.93 mmol) was added and solution was purged with argon for 15 min. ThenPd(PPh₃)₄ (255 mg, 0.22 mmol) was added and argon was purged again for15 min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the desired compound (800 mg,98%).

Step 3: Synthesis of5-(ethylamino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxylicacid

Aqueous NaOH (130 mg, 3.25 mmol) was added to a stirred solution ofcompound 6 (800 mg, 2.17 mmol) in ethanol (10 mL) and stirring continuedat 60° C. for 1 h. After completion, ethanol was removed under reducedpressure and acidified using dilute HCl up to pH-6. Aqueous layer wasextracted ethyl acetate (5 times) and the combined organic layer wasdried over sodium sulphate. Removal of the solvent under reducedpressure afforded the desired compound (700 mg, 91%). LCMS: 355.05(M+1)⁺; HPLC: 89.74% (@ 254 nm) (R_(t); 3.854; Method: Column: YMC ODS-A150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (CD₃OD, 400 MHz) δ 8.24 (s, 1H), 7.88 (s, 1H), 7.84 (d,2H, J=8 Hz), 7.71 (d, 2H, J=8.4 Hz), 4.45 (s, 2H), 4.06 (d, 2H, J=11.2Hz), 3.79 (t, 2H, J=12 Hz), 3.53 (q, 2H, J=7.2 Hz), 3.40-3.43 (m, 2H),3.22-3.31 (m, 2H), 2.66 (s, 3H), 1.45 (t, 3H, J=7.2 Hz).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethylamino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

5-(Ethylamino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxylicacid (300 mg, 0.84 mmol) was dissolved in DMSO (2 mL) and 3-(aminomethyl)-4, 6-dimethylpyridin-2(1H)-one (257 mg, 1.69 mmol) was added toit. After 15 minutes stirring at room temperature PyBOP (660 mg, 1.26mmol) was added to the reaction mixture and stirring was continued forovernight. After completion, the reaction mass was poured into ice,extracted with 10% MeOH/DCM (5 times). Combined organic layer was washedwith water and dried over sodium sulphate. Removal of the solvent underreduced pressure followed by solvent washings afforded the titlecompound (100 mg, 24%). LCMS: 489.20 (M+1)⁺; HPLC: 96.41% (@ 254 nm)(R_(t); 4.060; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30 OC; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (s, 1H), 9.90 (s, 1H), 8.06 (t, 1H), 7.73 (d, 2H, J=7.6 Hz), 7.55(d, 2H, J=7.2 Hz), 6.80 (d, 2H, J=7.6 Hz), 5.86 (s, 1H), 4.38 (s, 2H),4.27 (d, 2H, J=4 Hz), 3.95 (m, 2H), 3.62-3.65 (m, 2H), 3.28-3.31 (m,2H), 3.20-3.24 (m, 2H), 3.14-3.19 (m, 2H), 2.20 (s, 3H), 2.10 (s, 3H),2.06 (s, 3H), 1.21 (t, 3H, J=6.8 Hz).

Example 118: Synthesis5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-(hydroxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of tert-butyldimethyl(prop-2-yn-1-yloxy)silane

To an ice cooled stirred solution of prop-2-yn-1-ol (10.0 g, 178.3 mmol)and imidazole (18.2 mg, 267.5 mmol) in dichloroethane (500 mL), wasadded TBDMSC1 (40.24 g, 267.5 mmol) and stirring continued at 0° C. for1.5 h. On completion, saturated aqueous ammonium chloride solution wasadded to the reaction mixture and extracted with ethyl acetate (3times). Combined organic layer was dried over sodium sulphate. Removalof the solvent under reduced pressure followed by solvent washingsafforded the desired compound (20 g, 67%).

Step 2: Synthesis of 5-((tert-butyldimethylsilyl)oxy)pent-3-yn-2-one

To a stirred solution of tert-butyldimethyl(prop-2-yn-1-yloxy)silane(20.0 g, 116.9 mmol) in THF (400 mL) at −78° C. was added n-BuLi (90 mL,140.0 mmol) and the reaction mixture was allowed to attain roomtemperature with in 2 h. Then the reaction mixture was cooled to −78° C.and borontrifluoride etherate (18 mL, 140.0 mmol) was added. After 10minutes stirring acetic anhydride (15 mL, 153.0 mmol) was added and thereaction mixture was allowed to attain room temperature with in 2.5 h.Reaction was quenched with aqueous 1N NaOH solution and extracted withethyl acetate (3 times). Combined organic layer was dried over sodiumsulphate. Removal of the solvent under reduced pressure followed bysolvent washings afforded C (13 g, 52%).

Step 3: Synthesis of4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile

To a stirred solution of compound5-((tert-butyldimethylsilyl)oxy)pent-3-yn-2-one (13.0 g, 61.0 mmol) andcyanoacetamide (6.2 g, 73.2 mmol) in a mixture of ethanol and water(9:1) (270 mL) at room temperature was added piperidineacetate(catalytic) and the reaction mixture was heated to reflux for 5 h. Afterremoval of solvent water was added and the solid product was filtered.The solid product on washing with water followed by ether and hexaneafforded the desired compound (5.5 g, 32%).

Step 4: Synthesis of3-(aminomethyl)-4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methylpyridin-2(1H)-one

To a stirred solution of4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile(5.5 g, 19.7 mmol) in methanol (100 mL) and ammonia (30 mL) was addedRaney nickel (quantitative) and the reaction mixture was stirred inpresence of hydrogen under balloon pressure for 14 h. On completion,reaction mixture was filtered through celite and washed with methanol.Removal of the solvent under reduced pressure afforded the desiredcompound (3.5 g, 63%).

Step 5: Synthesis of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-(hydroxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

Aqueous NaOH (70 mg, 1.7 mmol) was added to a stirred solution of methyl5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate (400mg, 1.1 mmol) in ethanol (60 mL) and stirring continued at 60° C. for 1h. After completion, ethanol was removed under reduced pressure andacidified using dilute HCl up to pH ˜6. Aqueous layer was extractedethyl acetate (5 times) and the combined organic layer was dried oversodium sulphate. Removal of the solvent under reduced pressure affordedrespective acid (320 mg, 83.55%).

The above acid (400 mg, 1.1 mmol) was then dissolved in DMSO (4 mL) and3-(aminomethyl)-4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methylpyridin-2(1H)-one(525 mg, 1.7 mmol) was added to it. After 15 minutes stirring at roomtemperature PyBOP (900 mg, 1.6 mmol) was added to the reaction mixtureand stirring was continued for overnight. After completion, the reactionmass was poured into ice, extracted with 10% MeOH/DCM (5 times).Combined organic layer was washed with water and dried over sodiumsulphate. Removal of the solvent under reduced pressure followed bysolvent washings afforded the desired compound (230 mg, 40%).

Step 6: Synthesis5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-(hydroxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-(hydroxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(250 mg, 0.5 mmol) and4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (230mg, 7.6 mmol) in dioxane (5 mL), aqueous 2M Na₂CO₃ solution (0.9 mL, 1.8mmol) was added and solution was purged with argon for 15 min. Then Pd(PPh₃)₄ (57 mg, 0.05 mmol) was added and argon was purged again for 15min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the title compound (60 mg, 25%).LCMS: 589.35 (M+1)⁺; HPLC: 95.58% (@ 254 nm) (R_(t); 3.524; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 11.54 (s, 1H), 8.22(t, 1H), 7.57 (d, 2H, J=7.6 Hz), 7.38 (d, 2H, J=5.6 Hz), 7.36 (s, 1H),7.21 (s, 1H), 6.16 (s, 1H), 5.28 (m, 1H), 4.52 (d, 2H, J=4.8 Hz), 4.25(d, 2H, J=3.6 Hz), 3.81-3.83 (m, 2H), 3.57 (m, 4H), 3.48 (s, 2H),3.01-3.09 (m, 3H), 2.36 (m, 4H), 2.23 (s, 3H), 2.15 (s, 3H), 1.64-1.67(m, 2H), 1.51-1.53 (m, 2H), 1.23 (m, 2H), 0.82 (t, 3H, J=6.4 Hz).

Example 119: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-(N-(tetrahydro-2H-pyran-4-yl)acetamido)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-2-methyl-3-(N-(tetrahydro-2H-pyran-4-yl)acetamido) benzoate

A solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (520 mg,1.58 mmol) was heated at 70° C. in Acetic 3 ml of anhydride for 6 h. Thereaction mixture was cooled to room temperature and quenched with sat.NaHCO₃ and extracted with ethyl acetate. The organic layer was driedover Na₂SO₄, concentrated and purified by silica gel (100-200) columnchromatography to get the target compound (400 mg, 68%).

Step 2: Synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide

A mixture of methyl5-bromo-2-methyl-3-(N-(tetrahydro-2H-pyran-4-yl)acetamido) benzoate (400mg, 1.08 mmol) and NaOH (47 mg, 1.13 mmol) in 5 ml of ethanol:water(2:1) was heated at 70° C. for 2 h. The reaction mixture wasconcentrated to dryness and the crude material dissolved in water, pHwas adjusted to 5 to 6 by slow addition of HCl and extracted with 10%MeOH in DCM. The organic layer was dried over Na₂SO₄, concentrated underreduced pressure to afford 400 mg of acid.

The crude acid (400 mg, 1.23 mmol),3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (370 mg, 2.46 mmol), PyBOP(960 mg, 1.85 mmol) and triethyl amine (0.17 ml, 1.23) mixture wasstirred in 2 ml of DMSO at room temperature overnight. The reactionmixture was diluted with water and compound was extracted in 10% MeOH inDCM, dried over Na₂SO₄, concentrated and crude was purified by silicagel (100-200) column chromatography to get5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide(95 mg, 17.3%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-(N-(tetrahydro-2H-pyran-4-yl)acetamido)-[1,1′-biphenyl]-3-carboxamide

A solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-(N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide(50 mg, 0.10 mmol), (4-(morpholinomethyl)phenyl)boronic acid (41 mg,0.13 mmol), sodium carbonate (27 mg, 0.25 mmol) in 3 ml of dioxane wasdegassed with argon for 20 min, Pd(PPh₃) (12 mg, 0.0012 mmol) was addedto the mixture and heated to 100° C. for overnight. The reaction wascooled to room temperature and diluted with water, before extractionwith 10% MeOH in DCM, the organic layers were dried over Na₂SO₄,concentrated and the resulting crude product purified by silica gel(100-200) chromatography to obtain the title compound (26 mg, 23%).

LCMS: 609.35 (M+23)⁺; HPLC: 97.81% (@ 254 nm) (R_(t); 4.407; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 11.48 (bs, 1H), 8.41(t, 1H), 7.67-7.69 (m, 2H), 7.39-7.56 (m, 4H), 5.87 (s, 1H), 4.54-4.57(m, 1H), 4.30-4.31 (d, 2H, J=4 Hz), 3.77-3.85 (m, 2H), 3.50-3.58 (m,6H), 2.37 (m, 4H), 2.22 (s, 3H), 2.16 (s, 3H), 2.11 (s, 3H), 1.88-1.91(m, 1H), 1.51-1.65 (m, 6H), 2 protons merged in solvent peak.

Example 120: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-3′-fluoro-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-3′-fluoro-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(300 mg, 0.63 mmol) and (3-fluoro-4-formylphenyl)boronic acid (160 mg,0.94 mmol) in dioxane (6 mL), aqueous 2M Na₂CO₃ solution (1.15 mL, 2.3mmol) was added and solution was purged with argon for 15 min. ThenPd(PPh₃)₄ (72 mg, 0.06 mmol) was added and argon was purged again for 15min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the title compound (288 mg, 88%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-3′-fluoro-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-3′-fluoro-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide(285 mg, 0.55 mmol) and morpholine (149 mg, 1.64 mmol) in dichloroethane(5 mL), acetic acid (0.2 mL, 3.29 mmol) was added and reaction stirredat room temperature for 30 minutes. Then sodium triacetoxyborohydride(349 mg, 1.64 mmol) was added to the reaction mixture at 0° C., allowedto attain room temperature and stirring continued for overnight. Oncompletion, reaction mixture was diluted with dichloromethane, washedwith water, saturated aqueous sodium bicarbonate solution and dried oversodium sulphate. Removal of the solvent under reduced pressure followedby column chromatographic and prep. HPLC purification afforded the titlecompound (70 mg, 20%). LCMS: 591.45 (M+1)⁺; HPLC: 98.96% (@ 254 nm)(R_(t); 4.034; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.5 (bs, 1H), 10.1 (bs, 1H), 8.24 (s, 1H), 7.66-7.73 (m, 3H), 7.54 (s,1H), 7.36 (s, 1H), 5.88 (s, 1H), 4.44 (s, 2H), 4.30 (m, 5H), 3.96 (m,2H), 3.66-3.86 (m, 6H), 3.17-3.34 (m, 4H), 2.27 (s, 3H), 2.22 (s, 3H),2.11 (s, 3H), 1.57-1.67 (m, 4H), 0.84 (t, 3H, J=6 Hz).

Example 121: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2′-fluoro-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2′-fluoro-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(300 mg, 0.62 mmol) and (2-fluoro-4-formylphenyl)boronic acid (158 mg,0.94 mmol) in dioxane (3 mL), aqueous 2M Na₂CO₃ solution (1.13 mL, 2.26mmol) was added and solution was purged with argon for 15 min. ThenPd(PPh₃)₄ (72 mg, 0.06 mmol) was added and argon was purged again for 15min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the desired compound (300 mg,91%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2′-fluoro-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2′-fluoro-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide(300 mg, 0.57 mmol) and morpholine (100 mg, 1.15 mmol) in dichloroethane(4 mL), acetic acid (207 mg, 3.46 mmol) was added and reaction stirredat room temperature for 30 minutes. Then sodium triacetoxyborohydride(367 mg, 1.73 mmol) was added to the reaction mixture at 0° C., allowedto attain room temperature and stirring continued for overnight. Oncompletion, reaction mixture was diluted with dichloromethane, washedwith water, saturated aqueous sodium bicarbonate solution and dried oversodium sulphate. Removal of the solvent under reduced pressure followedby column chromatographic purification afforded the title compound (300mg, 87.97%).

LCMS: 591.30 (M+1)⁺; HPLC: 96.03% (@ 254 nm) (R_(t); 4.077; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs, 1H), 8.17(t, 1H), 7.47 (t, 1H, J=8 Hz), 7.30 (s, 1H), 7.21-7.23 (m, 2H), 7.10 (s,1H), 5.85 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.81-3.84 (m, 2H), 3.50-3.59(m, 6H), 3.22-3.25 (m, 2H), 3.00-3.06 (m, 3H), 2.38 (m, 4H), 2.25 (s,3H), 2.19 (s, 3H), 2.10 (s, 3H), 1.51-1.66 (m, 4H), 0.83 (t, 3H, J=6.8Hz).

Example 122: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2′,4-dimethyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4′-formyl-2′,4-dimethyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(400 mg, 0.84 mmol) and3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde(310 mg, 1.26 mmol) in dioxane (2 mL), aqueous 2M Na₂CO₃ solution (1.5mL, 3.03 mmol) was added and solution was purged with argon for 15 min.Then Pd(PPh₃)₄ (97 mg, 0.08 mmol) was added and argon was purged againfor 15 min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the desired compound (300 mg,69.28%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2′,4-dimethyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4′-formyl-2′,4-dimethyl-[1,1′-biphenyl]-3-carboxamide(410 mg, 0.79 mmol) and morpholine (210 mg, 2.38 mmol) in dichloroethane(10 mL), acetic acid (280 mg, 4.77 mmol) was added and reaction stirredat room temperature for 30 minutes. Then sodium triacetoxyborohydride(580 mg, 2.71 mmol) was added to the reaction mixture at 0° C., allowedto attain room temperature and stirring continued for overnight. Oncompletion, reaction mixture was diluted with dichloromethane, washedwith water, saturated aqueous sodium bicarbonate solution and dried oversodium sulphate. Removal of the solvent under reduced pressure followedby column chromatographic purification afforded the title compound (125mg, 26.76%).

LCMS: 587.55 (M+1)⁺; HPLC: 97.23% (@ 254 nm) (R_(t); 4.065; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs, 1H), 9.91(bs, 1H), 8.17 (bs, 1H), 7.32-7.42 (m, 3H), 7.15 (bs, 1H), 6.92 (bs,1H), 5.86 (s, 1H), 4.27-4.35 (m, 4H), 3.86 (m, 2H), 3.64-3.67 (m, 3H),3.12-3.32 (m, 10H), 2.33 (bs, 6H), 2.19 (s, 3H), 2.10 (s, 3H), 1.55-1.64(m, 4H), 0.84 (t, 3H, J=6 Hz), 2 protons merged in solvent peak.

Example 123: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholine-4-carbonyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(300 mg, 0.63 mmol) andmorpholino(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone(260 mg, 0.82 mmol) in dioxane (10 mL), aqueous 2M Na₂CO₃ solution (1.13mL, 2.27 mmol) was added and solution was purged with argon for 15 min.Then Pd(PPh₃)₄ (72 mg, 0.06 mmol) was added and argon was purged againfor 15 min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the title compound (250 mg, 68%).LCMS: 587.35 (M+1)⁺; HPLC: 93.85% (@ 254 nm) (R_(t); 4.535; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 11.46 (bs, 1H), 8.21(t, 1H), 7.69-7.71 (m, 2H), 7.45-7.49 (m, 3H), 7.26 (s, 1H), 5.86 (s,1H), 4.29 (d, 2H, J=4 Hz), 3.82-3.84 (m, 2H), 3.48-3.60 (m, 8H),3.23-3.25 (m, 2H), 3.09-3.11 (m, 3H), 2.26 (s, 3H), 2.21 (s, 3H), 2.11(s, 3H), 1.52-1.68 (m, 4H), 0.83 (t, 3H, J=6.8 Hz).

Example 124: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(1-(methylsulfonyl)piperidin-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(piperidin-4-yl)amino)-4-methyl-4′-(morpholinomethyly)-[1,1′-biphenyl]-3-carboxamide(0.2 g, 0.35 mmol) in DCM (8 mL) was added triethyl amine (0.106 g, 1.04mmol) and mesyl chloride (0.08 g, 0.69 mmol) at 0° C. The resultingreaction mixture was stirred at room temperature for 2 h. Aftercompletion, water was added to the reaction mixture and extracted with10% MeOH/DCM. Combined organic layers were dried over sodium sulphateand concentrated under reduced pressure giving crude material which wasthen dissolved in methanol (10 mL) and added NaOH (0.021 g, 0.52 mmol).This mixture was stirred at room temperature for 15 h. After completion,extraction was carried out using 20% MeOH/DCM. Combined organic layerswere dried over sodium sulfate, concentrated and crude material waspurified by solvent washing giving the title compound (0.1 g, 45.45%).

LCMS: 650.85 (M+1)⁺; HPLC: 95.37% (@ 254 nm) (R_(t); 4.258; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs, 1H), 8.18(t, 1H), 7.57-7.59 (m, 3H), 7.37-7.39 (m, 2H), 7.22 (s, 1H), 5.86 (s,1H), 4.29 (d, 2H, J=4.4 Hz), 3.58 (m, 4H), 3.48-3.52 (m, 4H), 3.09-3.11(m, 2H), 2.94 (m, 1H), 2.82 (s, 3H), 2.67-2.72 (m, 2H), 2.36 (m, 4H),2.24 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.81-1.83 (m, 2H), 1.59-1.61(m, 2H), 0.84 (t, 3H, J=6 Hz).

Example 125: Synthesis of5-((1-acetylpiperidin-4-yl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(piperidin-4-yl)amino)-4-methyl-4′-(morpholinomethyl))-[1,1′-biphenyl]-3-carboxamide(0.25 g, 0.44 mmol) and acetic acid (0.052 g, 0.86 mmol) in DMF (3 mL),EDCI (0.123 g, 0.64 mmol) and HOBt (0.087 g, 0.64 mmol) was addedfollowed by the addition of triethylamine (0.108 g, 1.06 mmol) andreaction was stirred at room temperature for overnight. After completionof the reaction, water was added to it and extraction was carried outusing 10% MeOH/DCM. The combined organic layers were washed with water,dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford crude material which was purified by columnchromatography to give the title compound (0.1 g, 37.31%).

LCMS: 614.75 (M+1)⁺; HPLC: 97.57% (@ 254 nm) (R_(t); 4.140; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs, 1H), 8.19(t, 1H), 7.57 (d, 2H, J=7.2 Hz), 7.37-7.39 (m, 3H), 7.22 (s, 1H), 5.86(s, 1H), 4.29 (d, 2H, J=4.4 Hz), 3.78 (m, 1H), 3.49-3.58 (m, 6H),2.99-3.08 (m, 4H), 2.36 (m, 4H), 2.24 (s, 3H), 2.21 (s, 3H), 2.10 (s,3H), 1.97 (s, 3H), 1.74 (m, 2H), 1.31-1.52 (m, 2H), 0.83 (t, 3H, J=6.8Hz), 2 protons merged in solvent peak.

Example 126: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(piperidin-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of tert-butyl4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl)amino)piperidine-1-carboxylate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (4.5 g,18.44 mmol) and tert-butyl 4-oxopiperidine-1-carboxylate (11.01 g, 55.33mmol) in dichloroethane (50 mL), acetic acid (6.64 g, 110.6 mmol) wasadded and reaction stirred at room temperature for 10 min. Then sodiumtriacetoxyborohydride (11.72 g, 55.28 mmol) was added at 0° C. andreaction stirred overnight at room temperature. On completion, solventwas removed under reduced pressure and crude material was purified bycolumn chromatography to afford the desired compound (5.2 g, 66.24%).

Step 2: Synthesis of tert-butyl4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl)(ethyl)-amino)-piperidine-1-carboxylate

To a stirred solution of tert-butyl4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl)amino)piperidine-1-carboxylate(5 g, 11.70 mmol) and acetaldehyde (1.58 g, 35.12 mmol) indichloroethane (60 mL), acetic acid (4.24 g, 70.66 mmol) was added andreaction stirred at room temperature for 10 min. Then sodiumtriacetoxyborohydride (7.44 g, 35.09 mmol) was added at 0° C. andreaction stirred at room temperature for 2 h. On completion, thereaction was quenched with aqueous sodium bicarbonate, organic phase wasseparated and aqueous phase was extracted with dichloromethane. Combinedorganic layers were dried over sodium sulphate and concentrated underreduced pressure to give crude material was purified by columnchromatography to afford the desired product (5 g, 93.45%).

Step 3: Synthesis of tert-butyl4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-phenyl)(ethyl)amino)piperidine-1-carboxylate

Aqueous NaOH (0.7 g, 17.50 mmol) was added to a solution of tert-butyl4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl)amino)piperidine-1-carboxylate(5 g, 10.94 mmol) in ethanol (50 mL) and stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and acidified using dilute HCl up to pH 6 and adjusted usingcitric acid to pH 4. Extraction was carried out using ethyl acetate.Combined organic layers were dried concentrated giving respective acid(4.8 g, 99.17%).

The above acid (4.8 g, 10.90 mmol) was then dissolved in DMSO (20 mL)and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (3.32 g, 21.81 mmol)was added to it. The reaction mixture was stirred at room temperaturefor 15 min before PYBOP (8.50 g, 16.35 mmol) was added to it andstirring was continued for overnight. On completion, reaction mixturewas diluted with water and extracted with 10% MeOH/DCM. Combined organiclayers were dried over Na₂SO₄ and solvent removed under reduced pressureto afford crude material which was purified by column chromatographyover silica gel to afford tert-butyl4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-phenyl)(ethyl)amino)piperidine-1-carboxylate(4.4 g, 70.96%).

Step 4: Synthesis of tert-butyl4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(ethyl)amino)piperidine-1-carboxylate

To a stirred solution of tert-butyl4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-phenyl)(ethyl)amino)piperidine-1-carboxylate(2 g, 3.47 mmol) and 4-(morpholinomethyl)-phenyl)boronic acid (1.58 g,5.21 mmol) in dioxane/water mixture, Na₂CO₃ (1.32 g, 12.45 mmol) wasadded and solution purged with argon for 15 min. Then Pd (PPh₃)₄ (0.4 g,0.35 mmol) was added and argon was purged again for 10 min. The reactionmixture was heated at 90° C. for 3.5 h. On completion, reaction mixturewas diluted with water and extracted with 10% MeOH/DCM. Combined organiclayers were dried over Na₂SO₄ and solvent removed under reduced pressureto afford crude material which was purified by column chromatographyover silica gel to afford desired compound (1.6 g, 68.66%).

Step 5: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(piperidin-4-yl)amino)-4-methyl-4′-(morpholinomethyly)-[1,1′-biphenyl]-3-carboxamide

tert-Butyl4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(ethyl)amino)piperidine-1-carboxylate(1.3 g, 0.1.93 mmol) was taken in DCM (20 mL), to it TFA (10 mL) wasadded at 0° C. and stirred at rt for 2 h. On completion of reaction,solvent was removed under reduced pressure, reaction was quenched withaqueous sodium bicarbonate and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over sodium sulphate and concentrated underreduced pressure to obtain crude; which then purified by acetonitrilewashing to give the title compound (0.9 g, 81.81%).

LCMS: 572.35 (M+1)⁺; HPLC: 96.59% (@ 254 nm) (R_(t); 3.964; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 8.19 (t, 1H), 7.57(d, 2H, J=8 Hz), 7.38 (m, 3H), 7.21 (s, 1H), 5.86 (s, 1H), 4.29 (d, 2H,J=4 Hz), 3.48-3.57 (m, 8H), 2.98-3.10 (m, 4H), 2.88 (m, 1H), 2.36 (m,4H), 2.24 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.70-1.73 (m, 2H),1.48-1.51 (m, 2H), 0.84 (t, 3H, J=6.8 Hz).

Example 127: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(1-pivaloylpiperidin-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(piperidin-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide(0.2 g, 0.34 mmol) was dissolved in DMSO (2 mL) and pivalic acid (0.107g, 1.04 mmol) and triethyl amine (0.106 g, 1.04 mmol) was added to it.The reaction mixture was stirred at room temperature for 15 min beforePYBOP (0.27 g, 0.52 mmol) was added to it and stirring was continued forovernight. After completion of the reaction, reaction mixture was pouredinto ice, extracted with 10% MeOH/DCM. Combined organic layers weredried, concentrated to obtain crude; which then purified by columnchromatography to afford the title compound (0.14 g, 60.86%).

LCMS: 656.65 (M+1)⁺; HPLC: 97.51% (@ 254 nm) (R_(t); 4.555; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs, 1H), 8.18(t, 1H), 7.57 (d, 2H, J=7.2), 7.37-7.40 (m, 3H), 7.21 (s, 1H), 5.86 (s,1H), 4.21-4.29 (m, 4H), 3.49-3.58 (m, 6H), 3.06-3.08 (m, 3H), 2.73-2.79(m, 2H), 2.37 (m, 4H), 2.24 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H),1.75-1.78 (m, 2H), 1.38-1.41 (m, 2H), 1.17 (s, 9H), 0.83 (t, 3H, J=7.2Hz).

Example 128: Synthesis of5-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5 g,20.57 mmol) and tert-butyl (4-oxocyclohexyl)carbamate (5.6 g, 26.74mmol) in dichloroethane (50 mL), acetic acid (7.4 g, 123.33 mmol) wasadded and reaction stirred at room temperature for 10 min. Then sodiumtriacetoxyborohydride (13 g, 61.72 mmol) was added at 0° C. and reactionstirred overnight at room temperature. On completion, the reaction wasquenched with aqueous sodium bicarbonate, organic phase was separatedand aqueous phase was extracted with dichloromethane. Combined organiclayers were dried over sodium sulphate and concentrated under reducedpressure to give crude material was purified by column chromatography toafford the title compound (3.5 g, 38.88%).

Step 2: Synthesis of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)cyclohexyl)-(ethyl)-amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate(1.4 g, 3.18 mmol) and acetaldehyde (0.419 g, 9.52 mmol) indichloroethane (20 mL), acetic acid (1.14 g, 19.0 mmol) was added andreaction stirred at room temperature for 10 min. Then sodiumtriacetoxyborohydride (2 g, 9.43 mmol) was added at 0° C. and reactionstirred at room temperature for 2 h. On completion, the reaction wasquenched with aqueous sodium bicarbonate, organic phase was separatedand aqueous phase was extracted with dichloromethane. Combined organiclayers were dried over sodium sulphate and concentrated under reducedpressure to give crude material was purified by column chromatography toafford the title compound (1.25 g, 84.45%).

Step 3: Synthesis of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate

Aqueous NaOH (0.16 g, 4.0 mmol) was added to a solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)cyclohexyl)-(ethyl)-amino)-2-methylbenzoate(1.25 g, 2.67 mmol) in ethanol (10 mL) and stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and acidified using dilute HCl up to pH 6 and adjusted usingcitric acid to pH 4. Extraction was carried out using ethyl acetate.Combined organic layers were dried concentrated giving respective acid(1.1 g, 90%).

The above acid (1.1 g, 2.42 mmol) was then dissolved in DMSO (10 mL) and3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.736 g, 4.84 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (1.88 g, 3.61 mmol) was added to it and stirring wascontinued for overnight. On completion, the reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford the title compound (0.75 g, 53.57%).

Step 4: Synthesis of tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)-(ethyl)-amino)-cyclohexyl)-carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate(0.7 g, 1.19 mmol) and (4-(morpholinomethyl)phenyl)-boronic acid (0.489g, 1.78 mmol) in dioxane/water mixture, Na₂CO₃ (0.454 g, 4.28 mmol) wasadded and solution purged with argon for 15 min. Then Pd (PPh₃)₄ (0.137g, 0.119 mmol) was added and argon was purged again for 10 min. Thereaction mixture was heated at 90° C. for 3.5 h. On completion, thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.Combined organic layers were dried over Na₂SO₄ and solvent removed underreduced pressure to afford crude material which was purified by columnchromatography over silica gel to afford the title compound (0.55 g,67.48%).

Step 5: Synthesis of5-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

tert-Butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)-(ethyl)-amino)-cyclohexyl)-carbamate(0.55 g, 0.80 mmol) was taken in DCM (10 mL), to it, TFA (3 mL) wasadded at 0° C. and stirred at room temperature overnight. On completionof reaction, solvent was removed under reduced pressure, and thereaction quenched with aqueous sodium bicarbonate and extracted withdichloromethane. Combined organic layers were dried over sodium sulphateand concentrated under reduced pressure to obtain crude product whichthen purified by acetonitrile washing to give the title compound (0.42g, 89.36%).

LCMS: 586.45 (M+1)⁺; HPLC: 98.38% (@ 254 nm) (R_(t); 3.667; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 8.18 (t, 1H), 7.56(d, 2H, J=7.6), 7.35-7.38 (m, 3H), 7.18 (s, 1H), 5.86 (s, 1H), 4.29 (d,2H, J=3.2 Hz), 3.58 (m, 4H), 3.49 (m, 2H), 3.09-3.10 (m, 2H), 2.63-2.66(m, 2H), 2.37 (m, 4H), 2.21 (s, 3H), 2.20 (s, 3H), 2.11 (s, 3H),1.75-1.78 (m, 4H), 1.40-1.43 (m, 2H), 1.05-1.08 (m, 2H), 0.83 (t, 3H,J=6.4 Hz).

Example 129: Synthesis of5-(((1r,4r)-4-acetamidocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide(0.25 g, 0.42 mmol) and acetic acid (0.151 g, 0.85 mmol) in DMF (3 mL),EDCI (0.123 g, 0.64 mmol) and HOBt (0.057 g, 0.42 mmol) was addedfollowed by the addition of triethylamine (0.064 g, 0.63 mmol) andreaction was stirred at room temperature overnight. After completion ofthe reaction, water was added to it and extraction was carried out using10% MeOH/DCM. The combined organic layers were washed with water, driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto afford crude material which was purified by column chromatography togive the title compound (0.11 g, 41.04%).

LCMS: 628.35 (M+1)⁺; HPLC: 98.79% (@ 254 nm) (R_(t); 3.902; Method:Column: YMC ODS-A 150 mm×4.6 mm×5 ta; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs, 1H), 8.18(t, 1H), 7.56-7.66 (m, 3H), 7.36-7.38 (m, 3H), 7.18 (s, 1H), 5.86 (s,1H), 4.29 (d, 2H, J=4 Hz), 3.99 (m, 1H), 3.48-3.58 (m, 6H), 3.10-3.11(m, 2H), 2.67 (m, 1H), 2.37 (m, 4H), 2.22 (s, 3H), 2.21 (s, 3H), 2.11(s, 3H), 1.74-1.79 (m, 6H), 1.43-1.46 (m, 2H), 1.08-1.11 (m, 2H),0.81-0.94 (t, 4H).

Example 130: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-fluoro-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: 6-fluoro-2-methyl-3-nitrobenzoic acid

A solution of 2-fluoro-6-methylbenzoic acid (2 g, 12.98 mmol) inconcentrated H₂SO₄ (15.77 ml, 295.85 mmol) was cooled to −5° C. in anacetone/ice bath in air. A mixture of concentrated nitric acid (1.08 ml,16.87 mmol) and concentrated H₂SO₄ (1 ml, 18.76 mmol) was added dropwiseto the reaction mixture at −5 to 0° C. over 15 minutes. The pale yellowreaction mixture was stirred at −5 to 0° C. for 30 minutes before beingpoured onto ice (100 g). The resulting precipitate was filtered anddissolved in EtOAc (50 ml) and the organic phase was washed withdeionized water (25 ml) followed by brine (25 ml). The organic phase wasdried over MgSO4, filtered and concentrated under reduced pressure togive 2 g (77%) of 6-fluoro-2-methyl-3-nitrobenzoic acid as a whitesolid. LC-MS 99%, 1.31 min (3 minute LC-MS method), m/z=198.0, ¹H NMR(500 MHz, Chloroform-d) δ ppm 8.04 (dd, J=9.1, 5.0 Hz, 1H), 7.16 (t,J=8.6 Hz, 1H), 2.63 (s, 3H).

Step 2: Synthesis of 3-bromo-2-fluoro-6-methyl-5-nitrobenzoic acid

To a solution of 6-fluoro-2-methyl-3-nitrobenzoic acid (100 mg, 0.5mmol) in concentrated H₂SO₄ (0.5 ml, 9.38 mmol) was added1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (79 mg, 0.28 mmol) atroom temperature and under nitrogen. The reaction mixture was stirredfor 6 hours during which time a precipitate formed. The reaction mixturewas added slowly to deionized water (3 ml) and the resulting precipitatewas filtered. The solid was washed with deionized water (2 ml) and airdried for 2 hours to give 123 mg (88%) of3-bromo-2-fluoro-6-methyl-5-nitrobenzoic acid as a pale yellow solid.LC-MS 94%, 1.61 min (3 minute LC-MS method), m/z=275.9/277.9 (ES−), ¹HNMR (500 MHz, Chloroform-d) δ ppm 8.25 (d, J=6.2 Hz, 1H), 2.58 (s, 3H).

Step 3: Synthesis of methyl 3-bromo-2-fluoro-6-methyl-5-nitrobenzoate

To a solution of 3-bromo-2-fluoro-6-methyl-5-nitrobenzoic acid (2.41 g,8.67 mmol) in N,N-Dimethylformamide (25 ml) at room temperature andunder nitrogen was added K₂CO₃ (2.4 g, 17.34 mmol) followed byiodomethane (0.7 ml, 11.27 mmol). The reaction mixture was stirred atroom temperature for 2 hours before being diluted with deionized water(100 ml) and extracted with EtOAc (3×50 ml). The combined organic phaseswere washed with saturated NaHCO₃(aq) (50 ml) and then dried over MgSO4,filtered and concentrated under reduced pressure. The residue waspurified twice by FCC (50 g silica, Isolute cartridge, gradient ofeluents; 98:2 Heptane:EtOAc to 9:1 Heptane:EtOAc) to give 2.43 g (89%)of methyl 3-bromo-2-fluoro-6-methyl-5-nitrobenzoate as a white solid.LC-MS 99%, 2.18 min (3 minute LC-MS method), m/z=no ionization, ¹H NMR(500 MHz, Chloroform-d) δ ppm 8.22 (d, J=6.2 Hz, 1H), 4.00 (s, 3H), 2.48(s, 3H).

Step 4: Synthesis of methyl 3-amino-5-bromo-6-fluoro-2-methylbenzoate

To a solution of methyl 3-bromo-2-fluoro-6-methyl-5-nitrobenzoate (2.43g, 8.32 mmol) in Methanol (80 ml) at room temperature was added ammoniumchloride (4.37 g, 83.2 mmol) followed by deionized Water (40 ml). Themixture was heated to 70° C. in air before the addition of iron (2.79 g,49.92 mmol). The reaction turned brown over the 2.5 hours it was stirredat 70° C. This mixture was allowed to cool to room temperature and wasfiltered through Kieselgel. The filter pad was washed with MeOH (80 ml)and the filtrate concentrated under reduced pressure. The residue wasdissolved in saturated NaHCO₃(aq) (50 ml) and EtOAc (150 ml). The phaseswere separated and the organic phase was washed with saturatedNaHCO₃(aq) (50 ml) before being dried over MgSO₄, filtered andconcentrated under reduced pressure. The residue was purified by FCC (50g silica, Isolute cartridge, gradient of eluents; 95:5 Heptane:EtOAc to7:3 Heptane:EtOAc) to give 2.23 g (95%, 77% corrected yields) of methyl3-amino-5-bromo-6-fluoro-2-methylbenzoate as a yellow oil. The materialwas taken through the next step without further purification. LC-MS 81%,1.87 min (3 minute LC-MS method), m/z=261.9/263.9, ¹H NMR (500 MHz,Chloroform-d) δ ppm 6.89 (d, J=6.0 Hz, 1H), 3.94 (s, 3H), 3.60 (s, 2H),2.08 (s, 3H).

Step 5: Synthesis of methyl3-bromo-2-fluoro-6-methyl-5-[(oxan-4-yl)amino]benzoate

To a solution of methyl 3-amino-5-bromo-6-fluoro-2-methylbenzoate (2.23g, 8.08 mmol) in 1,2-Dichloroethane (32 ml) at room temperature andunder nitrogen was added oxan-4-one (1.49 ml, 16.17 mmol) followed byacetic acid (2.78 ml, 48.5 mmol). This solution was stirred for 5minutes before the addition of sodium triacetoxyborohydride (5.14 g,24.25 mmol) at room temperature. After stirring for 5.5 hours there wasno unreacted starting material present by LCMS. Deionized water (32 ml)was added and the mixture was neutralized with solid NaHCO₃. The phaseswere separated and the aqueous layer was extracted with EtOAc (2×32 ml).The combined organic extracts were dried over MgSO4, filtered andconcentrated under reduced pressure. The residue was purified by FCC (50g silica, Isolute cartridge, gradient of eluents; 95:5 Heptane:EtOAc to6:4 Heptane:EtOAc) to give 2.3 g (82%) of methyl3-bromo-2-fluoro-6-methyl-5-[(oxan-4-yl)amino]benzoate as a off-whitesolid. LC-MS 99%, 2.13 min (3 minute LC-MS method), m/z=245.9/247.9, ¹HNMR (500 MHz, Chloroform-d) δ ppm 6.78 (d, J=5.9 Hz, 1H), 4.01 (dt,J=11.9, 3.4 Hz, 2H), 3.93 (s, 3H), 3.53 (td, J=11.7, 2.1 Hz, 2H),3.49-3.42 (m, 1H), 3.34 (s, 1H), 2.04 (s, 5H), 1.48 (qd, J=11.0, 4.2 Hz,2H).

Step 6: Synthesis of methyl3-bromo-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzoate

To a solution of methyl3-bromo-2-fluoro-6-methyl-5-[(oxan-4-yl)amino]benzoate (500 mg, 1.44mmol) in 1,2-Dichloroethane (15 ml) at room temperature and undernitrogen was added acetaldehyde (0.81 ml, 14.44 mmol) followed by aceticacid (0.5 ml, 8.67 mmol). This solution was stirred for 5 minutes beforethe addition of sodium triacetoxyborohydride (3.06 g, 14.44 mmol) atroom temperature. After stirring for 2 hours deionized water (20 ml) wasadded and the mixture was neutralized with solid NaHCO₃. The phases wereseparated and the aqueous layer was extracted with EtOAc (2×20 ml). Thecombined organic extracts were dried over MgSO₄, filtered andconcentrated under reduced pressure. The residue was purified by FCC (10g silica, Isolute cartridge, gradient of eluents; 95:5 Heptane:EtOAc to85:15 Heptane:EtOAc) to give 519 mg (96%) of methyl3-bromo-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzoate as a paleyellow oil that solidified upon standing. LC-MS 94%, 2.45 min (3 minuteLC-MS method), m/z=373.9/375.9, ¹H NMR (500 MHz, Chloroform-d) δ ppm7.33 (d, J=6.6 Hz, 1H), 3.95 (s, 5H), 3.32 (td, J=11.7, 2.1 Hz, 2H),3.00 (q, J=7.1 Hz, 2H), 2.88 (tt, J=10.9, 4.0 Hz, 1H), 2.25 (s, 3H),1.73-1.54 (m, 4H), 0.85 (t, J=7.1 Hz, 3H).

Step 7: Synthesis of3-bromo-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzoic acid

To a solution of methyl3-bromo-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzoate (519 mg,1.39 mmol) in Tetrahydrofuran (13 ml) and MeOH (4 ml) was added 4M NaOH(13.87 ml). The reaction mixture was stirred at 50° C. in air for 72hours. The reaction mixture was acidified to pH 2-3 with 6M HCl andextracted with DCM (5×15 ml). The combined organic extracts were driedover MgSO₄, filtered and concentrated under reduced pressure to give 526mg (95%) of 3-bromo-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzoicacid as a beige foam. LC-MS 88%, 1.77 min (3 minute LC-MS method),m/z=359.9/361.9, ¹H NMR (500 MHz, Chloroform-d) δ ppm 7.43-7.31 (m, 1H),4.00 (d, J=11.3 Hz, 2H), 3.41-3.29 (m, 2H), 3.16-2.91 (m, 3H), 2.40 (s,3H), 1.84-1.59 (m, 4H), 0.99-0.82 (m, 3H).

Step 8: Synthesis of3-bromo-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzamide

To a solution of3-bromo-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzoic acid (200 mg,0.56 mmol) in N,N-Dimethylformamide (2 ml) at room temperature and undernitrogen was added PyBOP (346.72 mg, 0.67 mmol) followed byN-ethyl-N-(propan-2-yl)propan-2-amine (145 l, 0.83 mmol) and3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 104 mg, 0.61mmol). After stirring for 1 hour at room temperature no startingmaterial was observed by LCMS. EtOAc (20 ml) was added to the reactionmixture and this was then washed with deionized water (5 ml) followed bysaturated NaHCO₃(aq) (3×5 ml). The organic phase was dried over MgSO4,filtered and concentrated under reduced pressure. The residue was thenpurified by FCC (5 g silica, Isolute cartridge, gradient of eluents;100% DCM to 97:3 DCM:MeOH) to give 112 mg (41%) of3-bromo-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzamideas a pale yellow solid. LC-MS 97%, 1.85 min (3 minute LC-MS method),m/z=494.0/496.0, ¹H NMR (500 MHz, Chloroform-d) δ ppm 11.66 (s, 1H),7.23 (d, J=6.5 Hz, 1H), 5.95 (s, 1H), 4.65-4.43 (m, 2H), 3.93 (d, J=11.0Hz, 2H), 3.38-3.22 (m, 2H), 2.97 (q, J=7.0 Hz, 2H), 2.91-2.79 (m, 1H),2.37 (s, 3H), 2.24-2.11 (m, 6H), 1.72-1.53 (m, 4H), 0.83 (t, J=7.0 Hz,3H).

Step 9: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-fluoro-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a solution of3-bromo-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[ethyl(oxan-4-yl)amino]-2-fluoro-6-methylbenzamide(112 mg, 0.23 mmol) in 1,4-Dioxane (2 ml) and Water (1 ml) was added4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (103mg, 0.34 mmol) followed by Na₂CO₃ (84.04 mg, 0.79 mmol). The solutionwas purged with nitrogen for 5 minutes before the addition ofpalladium-triphenylphosphine (1:4) (26 mg, 0.02 mmol). The yellowmixture was then purged with nitrogen for 5 minutes before being heatedto 100° C. After 4 hours LCMS indicated no unreacted starting materialwas present. The darken reaction mixture was diluted with deionizedwater (5 ml) and extracted with 10% MeOH in DCM (5×5 ml). The combinedorganic extracts were dried over MgSO4, filtered and concentrated underreduced pressure. The residue was purified by FCC (5 g silica, Isolutecartridge, gradient of eluents; 99:1 DCM:MeOH to 95:5 DCM:MeOH) to give69 mg (52%) of the title compound as an off-white solid. LC-MS 97%, 2.70min (7 minute LC-MS method), m/z=591.2, ¹H NMR (500 MHz, Chloroform-d) δppm 12.10 (br s, 1H), 7.53-7.30 (m, 4H), 7.13 (d, J=7.4 Hz, 1H), 7.07(br s, 1H), 5.88 (s, 1H), 4.55 (br s, 2H), 3.93 (d, J=11.2 Hz, 2H),3.73-3.69 (m, 4H), 3.52 (s, 2H), 3.30 (t, J=10.8 Hz, 2H), 3.02 (q, J=6.9Hz, 2H), 2.92 (ddd, J=14.6, 10.7, 3.7 Hz, 1H), 2.46 (s, 4H), 2.35 (s,3H), 2.26 (s, 3H), 2.12 (s, 3H), 1.79-1.43 (m, 4H), 0.86 (t, J=7.0 Hz,3H).

Example 131: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((3-oxomorpholino)methyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of4′-(bromomethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To an ice cooled stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4′-(hydroxymethyl)-4-methyl-[1,1′-biphenyl]-3-carboxamide(450 mg, 0.89 mmol) in DCM (10 mL) was added triphenyl phosphine (469mg, 1.78 mmol) and carbon tetrabromide (741 mg, 2.25 mmol). Reactionmixture was allowed to attain room temperature and stirring continuedfor 16 h. On completion, removal of the solvent under reduced pressurefollowed by column chromatographic purification afforded compound 8 (300mg, 59%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-((3-oxomorpholino)methyl)-[1,1′-biphenyl]-3-carboxamide

To an ice cooled stirred solution of4′-(bromomethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide(250 mg, 0.44 mmol) and morpholin-3-one (67 mg, 0.66 mmol) in DMF (30mL) was added sodium hydride (27 mg, 0.66 mmol). After 10 minutes, icewas removed and stirring continued for 16 h at room temperature. Oncompletion, water was added and extracted with DCM (3 times). Combinedorganic layer was dried over sodium sulphate. Removal of the solventunder reduced pressure followed by column chromatographic and prep. HPLCpurification afforded the title compound (75 mg, 29%). LCMS: 587.35(M+1)⁺; HPLC: 98.69% (@ 254 nm) (R_(t); 4.604; Method: Column: YMC ODS-A150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.50 (bs, 1H), 8.25 (m, 1H), 7.65(m, 2H), 7.37-7.35 (m, 3H), 5.87 (s, 1H), 4.59 (m, 2H), 4.29 (d, 2H),4.12 (s, 2H), 3.82 (m, 4H), 3.28 (m, 4H), 3.17-3.09 (m, 2H), 2.32-2.28(m, 4H), 2.22 (s, 3H), 2.11 (s, 3H), 1.57 (m, 4H), 0.86 (t, 3H).

Example 132: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-2-methyl-3-((tetrahydro-2H-thiopyran-4-yl)amino) benzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (2.5 g,10.24 mmol) and dihydro-2H-thiopyran-4(3H)-one (1.42 g, 12.29 mmol) indichloroethane (50 mL), acetic acid (3.6 ml, 61.47 mmol) was added andreaction stirred at room temperature for 10 min. The reaction mixturewas cooled to 0° C. and sodium triacetoxyborohydride (6.5 g, 30.73 mmol)was added and stirred at room temperature overnight. The reactionmixture was neutralized with sat. NaHCO₃ and compound was extracted inDCM, dried over Na₂SO₄, concentrated under reduced pressure. Columnchromatography purification of crude gave methyl5-bromo-2-methyl-3-((tetrahydro-2H-thiopyran-4-yl)amino) benzoate (2.5g, 71.0%).

Step 2: Synthesis of methyl5-bromo-3-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-thiopyran-4-yl)amino) benzoate (2.5g, 5.83 mmol) and acetaldehyde (513 mg, 11.66 mmol) in dichloroethane(50 mL), acetic acid (2.0 ml, 34.9 mmol) was added and reaction wasstirred at room temperature for 20 minutes. The reaction mixture wascooled to 0° C. and sodium triacetoxyborohydride (3.7 g, 17.49 mmol) wasadded and stirred at room temperature for overnight. The mixture wasneutralized with sat. NaHCO₃ and compound was extracted in DCM, driedover Na₂SO₄, concentrated under reduced pressure. The crude material waspurified by column chromatography to afford methyl5-bromo-3-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-2-methylbenzoate(2.0 g, 74.0%).

Step 3: Synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-2-methylbenzamide

A mixture of methyl5-bromo-3-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-2-methylbenzoate(2.0 g, 5.39 mol) and NaOH (0.323 g, 8.08 mol) in 3 ml of ethanol:water(2:1) was heated at 70° C. for 2 h. reaction mixture was concentrated todryness and crude was partitioned between water and DCM, organic layerwas dried over Na₂SO₄, concentrated under reduced pressure to afford 1.8g of acid.

The crude acid (1.8 g, 5.04 mmol),3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (1.53 mg, 10.08 mmol) andPyBOP (3.9 g, 7.56 mmol) mixture was stirred in 3 ml of DMSO at roomtemperature overnight. The reaction mixture was diluted with water andcompound was extracted in DCM. Organic layer was dried over Na₂SO₄,concentrated under reduced pressure and purified by silica gel (100-200mesh) column chromatography (eluted at 4% MeOH in DCM) to yield5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-2-methylbenzamide(1.5 g, 60.7%).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-2-methylbenzamide(800 mg, 1.629 mmol),4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine (740mg, 2.443 mmol), sodium carbonate (621 mg, 5.86 mmol) in 20 ml ofdioxane was degassed with argon for 20 min, Pd(PPh₃) (188 mg, 0.16 mmol)was added to the mixture and heated to 100° C. overnight. The reactionwas cooled to room temperature and diluted with water, compound wasextracted in 10% MeOH in DCM, dried over Na₂SO₄, concentrated and crudewas purified by silica gel (100-200) chromatography to obtain the titlecompound (700 mg, 73.0%).

LCMS: 589.25 (M+1)⁺; HPLC: 96.75% (@ 254 nm) (R_(t); 4.869; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (D20, 400 MHz) δ 7.78-7.89 (m, 4H),7.64-7.66 (m, 2H), 6.33 (s, 1H), 4.52 (s, 2H), 4.45 (s, 2H), 4.13 (d,J=13.2 Hz, 2H), 3.77-3.89 (m, 5H), 3.49 (d, J=12.0 Hz, 2H), 3.30-3.33(m, 2H), 2.73-2.82 (m, 5H), 2.44, 2.38, 2.30 (3s, 9H), 1.89 (m, 2H),1.06 (t, J=7.2 Hz, 3H).

Example 133: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(1-oxidotetrahydro-2H-thiopyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a cooled solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide(200 mg, 0.34 mmol) in 2 ml of DCM, m-CPBA (70 mg, 0.41 mmol) was addedat 0° C. and stirred at room temperature for 2 h (monitored by TLC). Thereaction was quenched with sat. NaHCO₃, compound was extracted in DCM,dried over Na₂SO₄, concentrated under reduced pressure and purified bysilica gel (100-200) column chromatography to obtain the title compound(60 mg, 29.3%).

LCMS: 605.25 (M+1)⁺; HPLC: 44.06% & 54.42% (@ 254 nm) (R_(t);4.092&4.448; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.45 (s, 1H), 8.18 (t, 1H), 7.59-7.57 (m, 2H), 7.39-7.37 (m, 3H),7.23-7.21 (m, 1H), 5.86 (s, 1H), 4.29 (d, 2H, J=4 Hz),3.58 (m, 3H), 3.48(m, 3H), 3.18-2.86 (m, 5H), 2.67-2.59 (m, 4H), 2.37-2.33 (m, 4H), 2.25(s, 3H), 2.21 (s, 3H), 2.10 (s, 3H), 1.77 (m, 2H), 0.85 (t, 3H).

Example 134:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-3′,4-dimethyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Compound 134 was prepared with the method similar to that described inExample 131. Analytical Data: LCMS: 587.4 (M+1)⁺; HPLC: 98.76% (@ 254nm) (R_(t); 4.11; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6, 400 MHz)δ 11.45 (bs, 1H), 8.17 (bs, 1H), 7.41-7.20 (m, 5H), 5.85 (s, 1H), 4.28(d, J=4 Hz, 2H), 3.82 (d, J=10 Hz, 2H), 3.55 (m, 4H), 3.44 (bs, 2H),3.27-3.22 (m, 2H), 3.09-3.01 (m, 3H), 2.39 (m, 7H), 2.23 (s, 3H), 2.20(s, 3H), 2.10 (s, 3H), 1.67-1.51 (m, 4H), 0.83 (t, J=6.8 Hz, 3H).

Example 135:4-((3′-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5′-(ethyl(1-oxidotetrahydro-2H-thiopyran-4-yl)amino)-4′-methyl-[1,1′-biphenyl]-4-yl)methyl)morpholine4-oxide

During above mentioned prep HPLC purification,4-((3′-(((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5′-(ethyl(1-oxidotetrahydro-2H-thiopyran-4-yl)amino)-4′-methyl-[1,1′-biphenyl]-4-yl)methyl)morpholine4-oxide was also isolated. LCMS: 621.40 (M+1)⁺; HPLC: 98.69% (@ 254 nm)(R_(t); 4.157; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ12.18 (s, 1H), 11.45 (s, 1H), 8.20 (t, 1H), 7.79 (d, 2H, J=6.8 Hz), 7.62(d, 2H, J=6.8 Hz), 7.45 (s, 1H), 7.27 (s, 2H), 5.86 (s, 1H), 4.89 (s,2H), 4.30 (d, 2H, J=4 Hz), 4.00-3.80 (m, 7H), 3.19 (m, 2H), 3.00-2.85(m, 4H), 2.70-2.60 (m, 2H), 2.30 (bs, 2H), 2.26 (s, 3H), 2.21 (s, 3H),2.10 (s, 3H), 1.75 (m, 2H), 0.87 (t, 3H, J=6 Hz).

Example 136: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)(ethyl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a cooled solution of compoundN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-thiopyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide(200 mg, 0.34 mmol) in 2 ml of DCM, m-CPBA (117 mg, 0.68 mmol) was addedat 0° C. and stirred at room temperature for 2 hours (monitored by TLC).The reaction was quenched with saturated NaHCO₃ and extracted with inDCM, dried over Na₂SO₄ and concentrated under reduced pressure. Aftercolumn chromatography, the title compound was obtained as the TFA saltafter further purification by prep. HPLC (80 mg, 38.1%).

LCMS: 621.45 (M+1)⁺; HPLC: 99.93% (@ 254 nm) (R_(t); 4.522; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 12.22 (s, 1H), 11.45(s, 1H), 8.20 (t, 1H), 7.78 (d, 2H, J=8 Hz), 7.61 (d, 2H, J=8 Hz), 7.43(s, 1H), 7.28 (s, 1H), 5.86 (s, 1H), 4.89 (s, 2H), 4.29 (d, 2H, J=4.4Hz), 4.00-3.80 (m, 7H), 3.32 (m, 2H), 3.04 (m, 4H), 2.65-2.55 (m, 2H),2.26 (s, 3H), 2.21 (s, 3H), 2.17 (m, 2H), 2.10 (s, 3H), 1.78 (m, 2H),0.83 (t, 3H, J=6.4&7.2 Hz).

Example 137: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4,4′-dimethyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4,4′-dimethyl-[1,1′-biphenyl]-3-carboxamide(200 mg, 0.35 mmol) in dichloromethane at room temperature, m-CPBA (60mg, 0.35 mmol) was added and stirring continued for overnight at roomtemperature. On completion, reaction was quenched by addition ofsaturated aqueous sodium bicarbonate solution and extracted with 10%MeOH/DCM (3 times). Combined organic layer was dried over sodiumsulphate. Removal of the solvent under reduced pressure followed bysolvent washing afforded the title compound (120 mg, 58%). LCMS: 589.35(M)+; HPLC: 95.56% (@ 254 nm) (R_(t); 4.143; Method: Column: YMC ODS-A150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 11.5 (bs, 1H), 8.22 (t, 1H),7.66-7.60 (m, 4H), 7.42 (s, 1H), 7.21 (s, 1H), 5.85 (s, 1H), 4.34-4.28(m, 4H), 4.12-4.07 (m, 2H), 3.83-3.81 (m, 2H), 3.62-3.60 (m, 2H),3.42-3.39 (m, 2H), 3.33-3.22 (m, 2H), 3.16-3.08 (m, 3H), 2.65-2.62 (m,2H), 2.25-2.10 (m, 9H), 1.67-1.51 (m, 4H), 0.83 (t, 3H, J=6.8 Hz).

Example 138: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-((tetrahydro-2H-pyran-4-yl)(2,2,2-trifluoroethyl)amino)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic acid

A solution of 5-bromo-2-methylbenzoic acid (5.0 g, 23 mmol) inconcentrated H₂SO₄ (27 ml, 512 mmol) was cooled to 5° C. in anacetone/ice bath. A mixture of concentrated nitric acid (1.9 ml, 30mmol) and concentrated H₂SO₄ (2.8 ml, 52 mmol) was added dropwise to thereaction mixture at −5 to 0° C. over 15 minutes. The yellow reactionmixture was stirred at −5 to 0° C. for 2 hours during which time ayellow precipitate formed. The reaction mixture was poured onto ice (150g) and the precipitate was then collected by filtration. The precipitatewas air dried to give the title compound (5.5 g, 52%) as a pale yellowsolid. LC-MS 57%, 1.82 min (3.5 minute LC-MS method), no ionization; ¹HNMR (500 MHz, DMSO-d₆) δ ppm 8.29 (s, 1H) 8.13 (d, J=1.58 Hz, 1H) 2.43(s, 3H).

Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate

To a solution of 5-bromo-2-methyl-3-nitrobenzoic acid (5.5 g, 21 mmol)in DMF (42 ml) under nitrogen, was added Na₂CO₃ (3.4 g, 32 mmol)followed by iodomethane (2.0 ml, 32 mmol). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted withdeionized water (150 ml) and extracted with EtOAc (4×50 ml). Thecombined organic phases were washed with saturated NaHCO₃ (aq) (2×50ml), dried over MgSO4, filtered and concentrated in-vacuum to give thetitle compound (6.3 g, 61%) as a yellow oil. LC-MS 57%, 2.20 min (3.5minute LC-MS method), no ionization; ¹H NMR (500 MHz, Chloroform-d) δppm 7.38 (d, J=2.05 Hz, 1H) 7.23 (d, J=2.05 Hz, 1H) 3.20 (s, 3H) 1.82(s, 3H).

Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate

To a solution of methyl 5-bromo-2-methyl-3-nitrobenzoate (6.3 g, 21mmol) in methanol (150 ml) was added ammonium chloride (11.0 g, 209mmol) followed by deionized water (75 ml). The mixture was heated to 70°C. before the addition of iron (7.0 g, 125 mmol). The reaction mixturewas stirred at 70° C. for 2 hours, before being allowed to cool to roomtemperature and filtered through Kieselgel. The filter pad was washedwith MeOH (150 ml) and the filtrate concentrated in-vacuo. The residuewas dissolved in saturated NaHCO₃ (aq) (50 ml) and EtOAc (150 ml). Thephases were separated and the organic phase was washed with saturatedNaHCO₃ (aq) (3×50 ml), dried over MgSO₄, filtered and concentratedin-vacuo. The residue was purified by flash column chromatography (50 gsilica Isolute cartridge, 5-20% EtOAc:Heptanes) to give the titlecompound (3.0 g, 51%) as a thick pale yellow oil. LC-MS 87%, 1.89 min(3.5 minute LC-MS method), m/z=243.9, 244.9, 245.9, 246.9; ¹H NMR (500MHz, Chloroform-d) δ ppm 7.34 (d, J=1.89 Hz, 1H) 6.95 (d, J=1.89 Hz, 1H)3.88 (s, 3H) 3.80 (br. s., 2H) 2.29 (s, 3H).

Step 4: Synthesis of methyl5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate

To a solution of methyl 3-amino-5-bromo-2-methylbenzoate (3.0 g, 12mmol) in 1,2-Dichloroethane (48 ml) under nitrogen, was added oxan-4-one(2.3 ml, 25 mmol) followed by acetic acid (4.2 ml, 74 mmol). Thereaction mixture was stirred for 5 minutes before the addition of sodiumtriacetoxyborohydride (7.8 g, 37 mmol). After stirring for 64 hours,deionized water (100 ml) was added and the mixture was neutralized withsolid NaHCO₃. The phases were separated and the aqueous layer wasextracted with EtOAc (4×50 ml). The combined organic extracts were driedover MgSO₄, filtered and concentrated in-vacuo. The residue was purifiedby flash column chromatography (50 g silica, Isolute cartridge, 10-30%EtOAc:Heptanes) to give the title compound (3.5 g 85%) as a white solid.LC-MS 99.8%, 2.18 min (3.5 minute LC-MS method), m/z=327.9, 328.9,329.9, 330.9; ¹H NMR (500 MHz, Chloroform-d) δ ppm 7.24 (d, J=1.73 Hz,1H) 6.85 (d, J=1.58 Hz, 1H) 4.03 (dt, J=11.82, 3.31 Hz, 2H) 3.88 (s, 3H)3.66 (br.s., 1H) 3.56 (td, J=11.55, 1.97 Hz, 2H) 3.47-3.55 (m, 1H) 2.24(s, 3H) 2.06 (d, J=13.56 Hz, 2H) 1.47-1.60 (m, 2H).

Step 5: Synthesis of methyl5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoate

In a 2 necked 100 ml RBF, containing methyl5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate (500 mg, 1.5 mmol) and TFA(15 ml), was added sodium tetrahydroborate (1.0 g, 26 mmol) portionwiseover 5 minutes. The reaction mixture was stirred at room temperature for2 hours and then heated to 50° C. for 3 hours and treated with a furtheraliquot of NaBH₄ (300 mg) over 25 minutes. The reaction mixture was thenheated to 60° C. for 2 hours and left to stir at room temperature for 17hours. The reaction mixture was treated with TFA (5 ml) and NaBH₄ (200mg) and heated back up to 60° C. for 3.5 hours. A further aliquot ofNaBH₄ (200 mg) was added over 15 minutes, along with TFA (5 ml) andheating continued for a further 3 hours, before being left to stand atroom temperature overnight. The reaction mixture was poured over ice (75ml) and stirred until the ice had melted. The reaction mixture was thenbasified by the addition of 6M NaOH (aq) (40 ml) and re-adjusted to pH 7using 1M HCl (aq) (40 ml). The resulting white suspension was collectedby filtration, the solid washed with water (20 ml) and dried in-vacuo at40° C. for 3 hours to give the title compound (577 mg, 91%) as a whitesolid. LC-MS 98.2%, 2.42 min (3.5 minute LC-MS method), m/z=409.90,410.9, 411.90, 412.9; ¹H NMR (500 MHz, Chloroform-d) δ ppm 7.80 (d,J=1.73 Hz, 1H) 7.41 (d, J=1.73 Hz, 1H) 4.01 (dd, J=11.51, 4.10 Hz, 2H)3.91 (s, 3H) 3.64 (d, J=5.20 Hz, 2H) 3.32 (t, J=11.82 Hz, 2H) 2.99 (tt,J=11.43, 3.63 Hz, 1H) 2.48 (s, 3H) 1.80 (dd, J=12.53, 1.50 Hz, 2H)1.54-1.62 (m, 2H).

Step 6: Synthesis of5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoic acid

To a stirred solution of methyl5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoate (572mg, 1.4 mmol) in a mixture of THF (14 ml) and MeOH (2.1 ml), was added4M NaOH (aq) (13.9 ml). The reaction mixture was stirred at 50° C. for5.5 hours and then stirred at room temperature for 17 hours. THF wasremoved by concentrating in-vacuo and the aqueous residue was acidifiedto pH 4 with 6M HCl (aq) (9.5 ml). The resulting suspension was allowedto stand at room temperature for 20 minutes before collecting the solidby filtration. The solid cake was washed with water (20 ml) and driedunder high vacuum for 2 hours to give the title compound (507 mg, 90%)as a white solid. LC-MS 98%, 2.04 min (3.5 minute LC-MS method),m/z=395.9, 396.9, 397.9, 398.9; ¹H NMR (500 MHz, Chloroform-d) δ ppm7.97 (d, J=1.73 Hz, 1H) 7.48 (d, J=1.73 Hz, 1H) 4.02 (dd, J=11.35, 3.94Hz, 2H) 3.65 (br. s, 2H) 3.33 (t, J=11.59 Hz, 2H) 3.00 (tt, J=11.49,3.80 Hz, 1H) 2.55 (s, 3H) 1.82 (d, J=11.98 Hz, 2H) 1.55-1.69 (m, 2H). OHnot visible.

Step 7: Synthesis of5-bromo-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzamide

A stirred solution of methyl5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoate (250mg, 0.63 mmol) in dry DMF (3.0 ml) at 0° C. under a balloon of nitrogen,was treated with HATU (288 mg, 0.76 mmol) and DIPEA (220 l, 1.3 mmol)dropwise. The resulting solution was stirred for 5 minutes and thentreated with 3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%,119 mg, 0.69 mmol). The resulting suspension was stirred at 0° C. for 20minutes and then stirred at room temperature for 16.5 hours. Thereaction mixture was treated with3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (30 mg). Stirringwas continued for further 23 hours and the reaction mixture was thenpartitioned between water (30 ml) and CH₂Cl₂ (20 ml). The layers wereseparated and the aqueous phase was extracted with CH₂Cl₂ (3×20 ml). Thecombined organics were washed with a saturated solution of NaHCO₃ (aq)(50 ml), water (60 ml), brine (2×40 ml), dried over MgSO₄, filtered andconcentrated in-vacuo. The crude residue was purified by flash columnchromatography (10 g SNAP cartridge, Isolera, 0-10% MeOH/CH₂Cl₂) andtriturated from ether (10 ml) with sonication. The resulting precipitatewas collected by filtration and dried in-vacuo to give the titlecompound (249 mg, 74%) as a white solid. LC-MS 100%, 4.08 min (7 minuteLC-MS method), m/z=530.0, 531.0, 532.0, 533.0; ¹H NMR (500 MHz, Acetone)δ 10.67 (s, 1H), 7.55 (d, J=1.8 Hz, 2H), 7.27 (d, J=1.9 Hz, 1H), 5.90(s, 1H), 4.40 (d, J=5.5 Hz, 2H), 3.90 (dd, J=11.2, 4.6 Hz, 4H), 3.28 (t,J=11.6 Hz, 2H), 3.07-2.97 (m, 1H), 2.32 (s, 3H), 2.29 (s, 3H), 2.24 (s,3H), 1.76 (dd, J=12.3, 1.6 Hz, 2H), 1.61 (qd, J=12.0, 4.5 Hz, 2H).

Step 8: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-5-((tetrahydro-2H-pyran-4-yl)(2,2,2-trifluoroethyl)amino)-[1,1′-biphenyl]-3-carboxamide

In a 2 necked RBF,5-bromo-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzamide(200 mg, 0.38 mmol),4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (126mg, 0.41 mmol) in 1,4-dioxane (3.0 ml) was treated with a solutioncontaining Na₂CO₃ (140 mg, 1.3 mmol) in water (1.0 ml). Nitrogen wasbubbled through the mixture using a long needle for 5 minutes before theaddition of palladium-triphenylphosphane (1:4) (44 mg, 0.04 mmol).Nitrogen was bubbled through the yellow suspension for a further 5minutes before heating the reaction mixture to 100° C. for 5.5 hours.The reaction mixture was diluted with water (10 ml) and 10% MeOH inCH₂Cl₂ (10 ml). The layers were separated and the aqueous phase wasextracted with 10% MeOH in CH₂Cl₂ (3×15 ml). The combined organicextracts were washed with brine (40 ml), dried over MgSO₄, filtered andconcentrated in-vacuo. The crude residue was purified by columnchromatography (10 g SNAP cartridge, Isolera, 0-4% MeOH:CH₂Cl₂) to givethe title compound (193 mg, 82%) as an off white powder. LC-MS 100%,3.34 min (7 minute LC-MS method), m/z=627.5; ¹H NMR (500 MHz, Acetone) δ10.76 (s, 1H), 7.65 (d, J=1.5 Hz, 1H), 7.60 (d, J=8.2 Hz, 2H), 7.57 (t,J=5.6 Hz, 1H), 7.43 (d, J=1.5 Hz, 1H), 7.39 (d, J=8.1 Hz, 2H), 5.91 (s,1H), 4.44 (d, J=5.5 Hz, 2H), 3.97 (s, 2H), 3.90 (dd, J=11.4, 4.1 Hz,2H), 3.61 (t, J=4.6 Hz, 4H), 3.50 (s, 2H), 3.29 (t, J=11.5 Hz, 2H), 3.06(tt, J=11.4, 3.8 Hz, 1H), 2.39 (d, J=5.0 Hz, 7H), 2.34 (s, 3H), 2.22 (s,3H), 1.82 (dd, J=12.3, 1.7 Hz, 2H), 1.70-1.56 (m, 2H).

Example 139: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-ethyl-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-chloro-2-[2-(trimethylsilyl)ethynyl]benzoate

To a solution of methyl 2-bromo-5-chlorobenzoate (14.8 g, 59 mmol) inTEA (124 ml, 889.82 mmol) was added copper(I) iodide (338 mg, 1.78 mmol)and triphenylphosphine (778 mg, 2.97 mmol) at room temperature and undernitrogen. This mixture had nitrogen bubbled through it for 10 minutesbefore the addition of ethynyl(trimethyl)silane (12.45 ml, 89 mmol) andPd(OAc)₂ (266 mg, 1.19 mmol). The reaction mixture was stirred at 50° C.for 20 hours before being concentrated under reduced pressure. Theresidue was dissolved in deionized water (50 ml) and EtOAc (50 ml) andfiltered through Celite. The filter cake was washed with EtOAc (50 ml)before the phases were separated and the aqueous layer was extractedwith EtOAc (2×50 ml). The combined organic extracts were dried overMgSO4, filtered and concentrated under reduced pressure. The residue waspurified by FCC (10 g silica, Isolute cartridge, gradient of eluents;99:1 Heptane:EtOAc to 85:15 Heptane:EtOAc) to give 16.2 g (102.4%) ofmethyl 5-chloro-2-[2-(trimethylsilyl)ethynyl]benzoate as an orange oilthat solidified upon standing. Sample contained heptane. LC-MS 91%, 2.57min (3 minute LC-MS method), m/z=267.4/268.9, ¹H NMR (500 MHz,Chloroform-d) δ ppm 7.89 (d, J=2.2 Hz, 1H), 7.51 (d, J=8.3 Hz, 1H), 7.41(dd, J=8.3, 2.3 Hz, 1H), 3.92 (s, 3H), 0.27 (s, 9H).

Step 2: Synthesis of methyl 5-chloro-2-ethynylbenzoate

To a solution of methyl 5-chloro-2-[2-(trimethylsilyl)ethynyl]benzoate(10 g, 37.5 mmol) in Methanol (150 ml) was added K₂CO₃ (10.36 g, 75mmol) at room temperature and in air. The reaction mixture was stirredfor 1 hour before being concentrated under reduced pressure. The residuewas dissolved in deionized water (50 ml) and EtOAc (50 ml). The phaseswere separated and the aqueous layer was extracted with EtOAc (2×50 ml).The combined organic extracts were dried over MgSO4, filtered andconcentrated under reduced pressure. The residue was purified by FCC (50g silica, Isolute cartridge, gradient of eluents; 95:5 Heptane:EtOAc to9:1 Heptane:EtOAc) to give 5.75 g (55.2%) of methyl5-chloro-2-ethynylbenzoate as an orange oil that solidified uponstanding. This material contained 30% of ethyl ester which was suitablefor use without any further purification. LC-MS 38%, 1.98 min (3 minuteLC-MS method), m/z=195.0/196.9, ¹H NMR (500 MHz, Chloroform-d) δ ppm7.93 (d, J=2.2 Hz, 1H), 7.55 (d, J=8.3 Hz, 1H), 7.45 (dd, J=8.3, 2.3 Hz,1H), 3.94 (s, 3H), 3.43 (s, 1H).

Step 3: Synthesis of methyl 5-chloro-2-ethylbenzoate

To a solution of methyl 5-chloro-2-ethynylbenzoate (5.34 g, 27.44 mmol)in Ethyl acetate (135 ml) was added Pd/C (10%) (50% water, 2.92 g, 1.37mmol). The reaction mixture was stirred under a hydrogen atmosphere atroom temperature for 3 hours. LCMS indicated the reaction had gone tocompletion and the mixture was filtered through Celite. The filter cakewashed with EtOAc (50 ml) and the filtrate was concentrated underreduced pressure to give 5.12 g (93.9%) of methyl5-chloro-2-ethylbenzoate as a brown oil which was suitable for usewithout any further purification. LC-MS 56%, 2.21 min (3 minute LC-MSmethod), m/z=198.9/200.9, ¹H NMR (500 MHz, Chloroform-d) δ ppm 7.84 (d,J=2.3 Hz, 1H), 7.39 (dd, J=8.3, 2.3 Hz, 1H), 7.21 (d, J=8.3 Hz, 1H),3.90 (s, 3H), 2.94 (q, J=7.5 Hz, 2H), 1.21 (t, J=7.5 Hz, 3H).

Step 4: Synthesis of methyl 5-chloro-2-ethyl-3-nitrobenzoate

A solution of methyl 5-chloro-2-ethylbenzoate (5.12 g, 25.77 mmol) inconcentrated H₂SO₄ (31 ml, 587 mmol) was cooled to −5° C. in anacetone/ice bath in air. A mixture of concentrated nitric acid (2.15 ml,33.51 mmol) and concentrated H₂SO₄ (2 ml, 37.52 mmol) was added dropwiseto the reaction mixture at −5 to 0° C. over 15 minutes. The pale yellowreaction mixture was stirred at −5 to 0° C. for 1 hour before beingpoured onto ice (500 ml) and this was extracted with EtOAc (3×100 ml).The combined organic phases was washed with deionized water (100 ml) andthen brine (100 ml). The organic phase was dried over MgSO4, filteredand concentrated under reduced pressure. LCMS and NMR showed ˜30%hydrolysis of the ester. The crude material was dissolved in Methanol(30 ml) and cooled to 0° C. under nitrogen where SOCl₂ (2.25 ml, 30.93mmol) was added slowly. The reaction mixture was then heated to refluxfor 6 hours before being concentrated under reduced pressure to give6.18 g (98.4%) of methyl 5-chloro-2-ethyl-3-nitrobenzoate as an orangeoil. Product contained 1:1 mixture of 3-nitro:6-nitro isomers along withsome ethyl ester which was suitable for use without any furtherpurification.

Step 5: Synthesis of methyl 3-amino-5-chloro-2-ethylbenzoate

To a solution of methyl 5-chloro-2-ethyl-3-nitrobenzoate (6.18 g, 25.36mmol) in Methanol (250 ml) at room temperature was added ammoniumchloride (13.31 g, 253.65 mmol) followed by deionized Water (125 ml).The mixture was heated to 70° C. in air before the addition of iron (8.5g, 152.19 mmol). The reaction turned to a dark color over the 2.5 hoursit was stirred at 70° C. This mixture was allowed to cool to roomtemperature and was filtered through Kieselgel. The filter pad waswashed with MeOH (250 ml) and the filtrate concentrated under reducedpressure. The residue was dissolved in saturated NaHCO₃(aq) (50 ml) andEtOAc (150 ml). The phases were separated and the organic phase waswashed with saturated NaHCO₃(aq) (2×50 ml) before being dried overMgSO₄, filtered and concentrated under reduced pressure. The residue waspurified by FCC (50 g silica, Isolute cartridge, gradient of eluents;95:5 Heptane:EtOAc to 75:25 Heptane:EtOAc) to give 2.42 g (22%, 7%corrected yields) of methyl 3-amino-5-chloro-2-ethylbenzoate as a yellowoil. The product contains ˜25% ethyl ester and possible ˜15% 4-nitroproducts. The material was taken through the next step without furtherpurification. LC-MS 31%, 2.00 min (3 minute LC-MS method), m/z=295.0, ¹HNMR (500 MHz, Chloroform-d) δ ppm 7.17 (d, J=2.1 Hz, 1H), 6.79 (d, J=2.1Hz, 1H), 3.87 (s, 3H), 3.86-3.81 (m, 2H), 2.74 (q, J=7.5 Hz, 2H), 1.20(t, J=7.5 Hz, 3H).

Step 6: Synthesis of methyl5-chloro-2-ethyl-3-[(oxan-4-yl)amino]benzoate

To a solution of methyl 3-amino-5-chloro-2-ethylbenzoate (1.5 g, 7.02mmol) in 1,2-Dichloroethane (28 ml) at room temperature and undernitrogen was added oxan-4-one (1.3 ml, 14.04 mmol) followed by aceticacid (2.41 ml, 42.12 mmol). This solution was stirred for 5 minutesbefore the addition of sodium triacetoxyborohydride (4.46 g, 21.06 mmol)at room temperature. After stirring for 20 hours, deionized water (28ml) was added and the mixture was neutralized with solid NaHCO₃. Thephases were separated and the aqueous layer was extracted with EtOAc(2×28 ml). The combined organic extracts were dried over MgSO4, filteredand concentrated under reduced pressure. The residue was purified by FCC(50 g silica, Isolute cartridge, gradient of eluents; 95:5 Heptane:EtOActo 8:2 Heptane:EtOAc) to give 1.76 g (84%, 50% corrected yields) ofmethyl 5-chloro-2-ethyl-3-[(oxan-4-yl)amino]benzoate as a white solid.Product contains ˜25% ethyl ester. The material was taken through thenext step without further purification. LC-MS 60%, 2.27 min (3 minuteLC-MS method), m/z=298.0/300.0, ¹H NMR (500 MHz, Chloroform-d) δ ppm7.07 (d, J=2.0 Hz, 1H), 6.71 (d, J=1.9 Hz, 1H), 4.01 (dt, J=11.8, 3.4Hz, 2H), 3.87 (s, 3H), 3.82-3.76 (m, 1H), 3.64-3.47 (m, 3H), 2.79-2.63(m, 2H), 2.06 (d, J=13.2 Hz, 2H), 1.55-1.46 (m, 2H), 1.18 (t, J=7.5 Hz,3H).

Step 7: Synthesis of 5-chloro-2-ethyl-3-[ethyl(oxan-4-yl)amino]benzoicAcid

To a solution of methyl 5-chloro-2-ethyl-3-[(oxan-4-yl)amino]benzoate(350 mg, 1.18 mmol) in DCE (10 ml) at room temperature and undernitrogen was added acetaldehyde (0.66 ml, 11.75 mmol) followed by aceticacid (0.4 ml, 7.05 mmol). This solution was stirred for 5 minutes beforethe addition of sodium triacetoxyborohydride (2.49 g, 11.75 mmol) atroom temperature. After stirring for 23 h further acetaldehyde (0.66 ml,11.75 mmol) was added followed sodium triacetoxyborohydride (2.49 g,11.75 mmol). After stirring for a further 3 hours deionized water (15ml) was added and the mixture was neutralized with solid NaHCO₃. Thephases were separated and the aqueous layer was extracted with EtOAc(2×15 ml). The combined organic extracts were dried over MgSO4, filteredand concentrated under reduced pressure. The residue was purified by FCC(10 g silica, Isolute cartridge, gradient of eluents; 99:1 Heptane:EtOActo 85:15 Heptane:EtOAc) to afford the title compound (317 mg) as a 2:1mixture of methyl and ethyl esters that were used in the next stage.

To the mixture of esters was added THF (10 ml) and 4M NaOH (9.7 ml, 38.9mmol) and the reaction was stirred at 50° C. for 27 hours, after whichtime MeOH (5 ml) was added to the reaction mixture and this was stirredfor a further 21H at 50° C. The reaction mixture was acidified to pH 2-3with 6M HCl and extracted with DCM (5×10 ml). The combined organicextracts were dried over MgSO4, filtered and concentrated under reducedpressure to afford the title compound as orange crystals (289 mg, 79%over two steps). LC-MS 100%, 2.09 min (3.5 minute LC-MS method),m/z=312.0/314.0, 1H NMR (500 MHz, Chloroform-d) δ 7.73 (d, J=1.7 Hz,1H), 7.29 (d, J=1.8 Hz, 1H), 3.99 (d, J=11.0 Hz, 2H), 3.38-3.29 (m, 2H),3.20-3.03 (m, 4H), 3.02-2.91 (m, 1H), 1.78-1.61 (m, 4H), 1.13 (t, J=7.4Hz, 3H), 0.91 (t, J=7.0 Hz, 3H).

Step 8: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-ethyl-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4′-(mrpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a solution of 5-chloro-2-ethyl-3-[ethyl(oxan-4-yl)amino]benzoic acid(191 mg, 0.61 mmol) in DMF (3 ml) at 0° C. was added HATU (280 mg, 0.74mmol) followed by DIPEA (213 l, 1.26 mmol) and3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 115 mg, 0.67mmol). The reaction was stirred at room temperature for 3 h after whichthe reaction was poured onto deionized water (50 ml) and the resultantsolid was filtered and washed with water. The aqueous phase was washedwith DCM (3×50 ml), the combined organics were washed with brine (30 m),dried with MgSO4, filtered and evaporated to give an oil. The solid andoil were combined an purified using a 10 g isolate column eluting with0% to 3% MeOH in DCM and evaporated followed by purification using a 10g Isolute eluting with 0% to 3% MeOH in EtOAc to afford the titlecompound as an off-white solid (234 mg, 79%). LC-MS 92%, 1.78 min (3.5minute LC-MS method), m/z=446.2/448. To a stirred solution of5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-[ethyl(oxan-4-yl)amino]benzamide(117 mg, 0.26 mmol) in a degassed mixture of diglyme (4 ml) and MeOH (2ml) which was bubbled with nitrogen gas was added2′-(dicyclohexylphosphanyl)-N,N-dimethylbiphenyl-2-amine (21 mg, 0.05mmol), palladium diacetate (5.89 mg, 0.03 mmol), caesium fluoride (120mg, 0.79 mmol) and4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (119mg, 0.39 mmol). The nitrogen bubbling continued for 10 min and then thereaction was heated to 70° C. for 16 h after which time further4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (119mg, 0.39 mmol) was added and heating continued for 6 h. The reaction wasthen cooled to room temperature and filtered through Keiselguhr, thecake being washed with MeOH. Distilled water (20 ml) was added to thefiltrate which was then extracted with EtOAc (3×50 ml), the combinedorganics were then washed with brine (2×50 ml), dried with MgSO4,filtered and evaporated. The resultant residue was purified using a 25 gIsolute column eluting with a gradient of 0% to 10% MeOH in DCM toafford the title compound as a pale yellow solid (32 mg, 21%). LC-MS99%, 2.72 min (7 minute LC-MS method), m/z=294.3 (M+H/2), 1H NMR (500MHz, Acetone-d6) δ 10.80 (s, 1H), 7.60-7.54 (m, 3H), 7.51 (s, 1H), 7.39(d, J=7.9 Hz, 2H), 7.36 (d, J=1.4 Hz, 1H), 5.92 (s, 1H), 4.45 (d, J=5.6Hz, 2H), 3.88 (d, J=8.0 Hz, 2H), 3.66-3.56 (m, 4H), 3.51 (s, 2H), 3.29(t, J=11.2 Hz, 2H), 3.17 (q, J=7.0 Hz, 2H), 3.09 (t, J=11.2 Hz, 1H),2.98 (q, J=7.4 Hz, 2H), 2.41 (s, 4H), 2.36 (s, 3H), 2.22 (s, 3H), 1.76(d, J=11.2 Hz, 2H), 1.65-1.56 (m, 2H), 1.08 (t, J=7.4 Hz, 3H), 0.91 (t,J=7.0 Hz, 3H).

Example 140: Synthesis of3′-cyano-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of 5-bromo-2-(morpholinomethyl)benzonitrile

To a stirred solution of 5-bromo-2-formylbenzonitrile (200 mg, 0.95mmol) and morpholine (248 mg, 2.85 mmol) in dichloroethane (10 mL),acetic acid (342 mg, 5.7 mmol) was added and reaction stirred at roomtemperature for 30 minutes. Then sodium triacetoxyborohydride (604 mg,2.85 mmol) was added to the reaction mixture at 0° C., allowed to attainroom temperature and stirring continued for overnight. On completion,reaction mixture was diluted with dichloromethane, washed with water,saturated aqueous sodium bicarbonate solution and dried over sodiumsulphate. Removal of the solvent under reduced pressure followed bycolumn chromatographic purification afforded the desired compound (150mg, 56%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

To a stirred mixture of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(1.0 g, 2.1 mmol), bispinacolato diboron (2.67 g, 10.5 mmol) andpotassium acetate (610 mg, 6.31 mmol) in dioxane (10 mL) was purged withargon for 15 min. Then1,1′-bis(diphenylphosphino)ferrocene-palladium(II)dichloridedichloromethane complex (85 mg, 0.10 mmol) was added and argon waspurged again for 15 min. Reaction mass was heated at 80° C. for 7 h. Oncompletion, reaction mixture was diluted with water and extracted with10% MeOH/DCM (3 times). Combined organic layer was dried over sodiumsulphate. Removal of the solvent under reduced pressure followed bycolumn chromatographic purification afforded the desired compound (250mg, 27%).

Step 3: Synthesis of3′-cyano-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of 5-bromo-2-(morpholinomethyl)benzonitrile (190mg, 0.68 mmol) andN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide(200 mg, 0.45 mmol) in dioxane (6 mL), aqueous 2M Na₂CO₃ solution (0.81mL, 1.63 mmol) was added and solution was purged with argon for 15 min.Then Pd(PPh₃)₄ (52 mg, 0.04 mmol) was added and argon was purged againfor 15 min. Reaction mass was heated at 100° C. for 4 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM(3 times). Combined organic layer was dried over sodium sulphate.Removal of the solvent under reduced pressure followed by columnchromatographic purification afforded the title compound (16 mg, 6%).LCMS: 598.20 (M+1)⁺; HPLC: 89.15% (@ 254 nm) (R_(t); 4.039; Method:Column: YMC ODS-A 150 mm×4.6 mm×5 L; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (bs, 1H), 8.20(t, 1H), 8.15 (s, 1H), 7.94 (d, 1H, J=6.8 Hz), 7.63 (d, 1H, J=8.4 Hz),7.51 (s, 1H), 7.31 (s, 1H), 5.86 (s, 1H), 4.29 (m, 2H), 3.84-3.82 (m,2H), 3.66-3.58 (m, 6H), 3.32 (m, 5H), 3.11-3.03 (m, 4H), 2.25 (s, 3H),2.21 (s, 3H), 2.11 (s, 3H), 1.65-1.51 (m, 4H), 0.82 (t, 3H, J=6 Hz).

Example 141:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid

To stirred solution of 2-methyl-3-nitrobenzoic acid (100 g, 552.48 mmol)in conc. H₂SO₄ (400 mL), 1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione(87.98 g, 307.70 mmol) was added in a portion-wise manner at roomtemperature. The reaction mixture was then stirred at room temperaturefor 5 h. The reaction mixture was poured into ice cold water, theprecipitated solid collected by filtration, washed with water and driedunder vacuum to afford desired 5-bromo-2-methyl-3-nitrobenzoic acid asoff-white solid (140 g, 97.90% yield). ¹H NMR (DMSO-d₆, 400 MHz) δ 8.31(s, 1H), 8.17 (s, 1H), 2.43 (s, 3H).

Step 2: methyl 5-bromo-2-methyl-3-nitrobenzoate

To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid (285 g,1104.65 mmol) in DMF (2.8 L) was added sodium carbonate (468 g, 4415.09mmol) followed by addition of methyl iodide (626.63 g, 4415 mmol) atroom temperature. The resulting reaction mixture was stirred at 60° C.for 8 h. The reaction mixture was then filtered to remove suspendedsolids which were washed well with ethyl acetate (3×1 L). The combinedfiltrates were washed well with water (5×3 L) and the aqueous phase backextracted with ethyl acetate (3×1 L). The combined organic extractsdried over anhydrous sodium sulfate, filtered and concentrated underreduced pressure to afford methyl 5-bromo-2-methyl-3-nitrobenzoate as anoff-white solid (290 g, 97% yield). ¹H NMR (CDCl₃, 400 MHz) δ 8.17 (s,1H), 7.91 (s, 1H), 3.96 (s, 3H), 2.59 (s, 3H).

Step 3: methyl 3-amino-5-bromo-2-methylbenzoate

To a stirred solution of methyl 5-bromo-2-methyl-3-nitrobenzoate (290 g,1058.39 mmol) in ethanol (1.5 L) was added aqueous ammonium chloride(283 g, 5290 mmol dissolved in 1.5 L water). The resulting mixture wasstirred and heated at 80° C. followed by addition of iron powder (472 g,8451 mmol) in portions at 80° C. The resulting reaction mixture washeated at 80° C. for 12 h. The reaction mixture was then hot filteredthrough Celite® and the Celite® bed washed well methanol (5 L) and thenwith 30% MeOH in DCM (5 L). The combined filtrates were concentrated invacuo and the residue obtained was diluted with aqueous bicarbonate (2L) and extracted with ethyl acetate (3×5 L). The combined organic layerswere dried over anhydrous sodium sulfate, filtered and concentratedunder reduced pressure to afford methyl 3-amino-5-bromo-2-methylbenzoateas a brown solid (220 g, 89.41% yield).

A portion of the product (5 g) was dissolved in hot ethanol (20 mL),insoluble residue filtered off and mother liquor concentrated to obtainmethyl 3-amino-5-bromo-2-methylbenzoate (3.5 g, 70% yield) with HPLCpurity 93.81% as light brown solid. ¹H NMR (CDCl₃, 400 MHz) δ 7.37 (s,1H), 6.92 (s, 1H), 3.94 (s, 3H), 3.80 (bs, 2H), 2.31 (s, 3H).

Step 4: methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5 g,20.5 mmol) and tert-butyl (4-oxocyclohexyl)carbamate (5.69 g, 26.7 mmol)in dichloroethane (50 mL), acetic acid (7.4 g, 123 mmol) was added andthe reaction was stirred at room temperature for 10 minutes. Sodiumtriacetoxyborohydride (13.1 g, 61.7 mmol) was then added at 0° C. andreaction was stirred at room temperature for 16 hours. The reaction wasquenched with aqueous sodium bicarbonate, the organic phase separatedand the aqueous phase extracted with dichloromethane. The combinedorganic layers were dried over anhydrous sodium sulfate and concentratedin vacuo. The crude product was purified by silica gel columnchromatography (100-200 mesh size) eluting with 10% ethyl acetate inhexane to afford 3.5 g of the more polar (trans) isomer, methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzoate,as solid (38.46%). ¹H NMR (CDCl₃, 400 MHz) δ 7.21 (s, 1H), 6.80 (s, 1H),4.41 (bs, 1H), 3.85 (s, 3H), 3.60 (m, 1H), 3.45 (m, 1H), 3.20 (m, 1H),2.22 (s, 3H), 2.15 (bs, 2H), 2.05 (bs, 2H), 1.45 (s, 9H), 1.30 (m, 4H).

Step 5: methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)-cyclohexyl)(ethyl)amino)-2-methylbenzoate(55 g, 0.124 mol) and acetaldehyde (11 g, 0.25 mol) in dichloroethane(550 mL), acetic acid (44.64 g, 0.744 mol) was added and the reactionmixture stirred at room temperature for 10 minutes. Sodiumtriacetoxyborohydride (79 g, 0.372 mol) was then added at 0° C. and thereaction mixture was stirred at room temperature for 16 hours. Thereaction was quenched with aqueous sodium bicarbonate, the organic phaseseparated and the aqueous phase extracted with dichloromethane. Thecombined extracts were dried over anhydrous sodium sulfate andconcentrated in-vacuo. The crude compound was purified by silica gelcolumn chromatography (100-200 mesh size) eluting with 10% ethyl acetatein hexane to afford 44 g of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-2-methylbenzoate(75.2%) as solid. ¹H NMR (DMSO-d₆, 400 MHz) δ 7.55 (s, 1H), 7.45 (s,1H), 6.65 (d, 1H), 3.80 (s, 3H), 3.15 (bs, 1H), 3.05 (q, 2H), 2.60 (m,1H), 2.30 (s, 3H), 1.75 (m, 4H), 1.40 (m, 2H), 1.35 (s, 9H), 1.10 (m,2H), 0.80 (t, 3H).

Step 6: tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate

Aqueous NaOH (3.5 g, 0.08 mol in 10 mL H₂O) was added to a solution ofmethyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-2-methylbenzoate(25 g, 0.053 mol) in EtOH (100 mL) and stirred at 60° C. for 1 h. Theethanol was then removed under reduced pressure and acidified to pH 8with dilute HCl and to pH 6 with citric acid. The mixture was extractedwith 10% methanol in DCM (3×200 mL). The combined organic layers weredried and concentrated giving the respective acid (24.2 g, 99.0%). ¹HNMR (DMSO-d₆, 400 MHz) δ 13.13 (s, 1H), 7.54 (s, 1H), 7.43 (s, 1H), 6.68(d, 1H), 3.14 (bs, 1H), 3.03 (q, 2H), 2.56 (m, 1H), 2.33 (s, 3H),1.80-1.65 (m, 4H), 1.40 (m, 2H), 1.35 (s, 9H), 1.10 (m, 2H), 0.77 (t,3H).

The acid (24 g, 0.053 mol) was dissolved in DMSO (100 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (16 g, 0.106 mol) andtriethylamine (5.3 g, 0.053 mol) was added. The reaction mixture wasstirred at room temperature for 15 min before PyBop (41 g, 0.079 mmol)was added and stirring was then continued for overnight at roomtemperature. The reaction mixture was poured into ice water (1 L). Theresulting precipitate was collected by filtration, washed well withwater (2×1 L) and dried. The product obtained was further purified bywashings with acetonitrile (3×200 mL) and DCM (100 mL) to affordtert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)-carbamate(24 g, 77%). ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47 (s, 1H), 8.24 (t, 1H),7.25 (s, 1H), 7.04 (s, 1H), 6.67 (d, 1H), 5.85 (s, 1H), 4.24 (d, 2H),3.13 (bs, 1H), 3.01 (q, 2H), 2.53 (m, 1H), 2.18 (s, 3H), 2.10 (s, 6H),1.80-1.65 (m, 4H), 1.40 (m, 2H), 1.35 (s, 9H), 1.10 (m, 2H), 0.77 (t,3H).

Step 7: tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(ethyl)amino)cyclohexyl)carbamate

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)-carbamate(24 g, 0.041 mol) and4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine (18g, 0.061 mol) in dioxane/water mixture (160 mL+40 mL), Na₂CO₃ (15 g,0.15 mol) was added and solution purged with argon for 15 min. Pd(PPh₃)₄(4.7 g, 0.041 mol) was then added and the reaction mixture again purgedwith argon for 10 min. The reaction mixture was heated at 100° C. for 4h. The reaction mixture was then diluted with 10% MeOH/DCM (500 mL) andfiltered. The filtrate was concentrated, diluted with water (500 mL) andextracted with 10% MeOH in DCM (3×500 mL). The combined organic layerswere dried over Na₂SO₄ and solvent removed under reduced pressure. Thecrude product was purified by silica gel column chromatography (100-200mesh) eluting with 7% MeOH in DCM to afford tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(ethyl)amino)cyclohexyl)carbamate(20 g, 71.43%). ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (s, 1H), 8.20 (t, 1H),7.56 (d, 2H), 7.36 (m, 3H), 7.17 (s, 1H), 6.66 (d, 1H), 5.85 (s, 1H),4.28 (d, 2H), 3.57 (bs, 4H), 3.48 (s, 2H), 3.20-3.05 (m, 3H), 2.62 (m,1H), 2.36 (bs, 4H), 2.20 (s, 6H), 2.10 (s, 3H), 1.75 (m, 4H), 1.42 (m,2H), 1.35 (s, 9H), 1.10 (m, 2H), 0.82 (t, 3H).

Step 8:5-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of tert-butyl((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-yl)(ethyl)amino)cyclohexyl)carbamate(20 g, 0.03 mol) in DCM (200 mL) at 0° C., TFA (75 mL) was added andreaction was stirred for 2 h at room temperature. The reaction mixturewas then concentrated to dryness and the residue basified with aqueoussaturated bicarbonate solution (300 mL) to pH 8. The mixture wasextracted with 20% methanol in DCM (4×200 m). The combined extracts weredried over Na₂SO₄ and the solvent removed under reduced pressure toafford5-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide(15.5 g, 91%) which was used as is in the next reaction. ¹H NMR(DMSO-d₆, 400 MHz) δ 8.18 (bs, 1H), 7.57 (d, 2H), 7.38 (m, 3H), 7.20 (s,1H), 5.85 (s, 1H), 4.29 (d, 2H), 3.57 (bs, 4H), 3.48 (s, 2H), 3.31 (bs,2H), 3.10 (m, 2H), 2.91 (m, 1H), 2.67 (m, 1H), 2.36 (bs, 4H), 2.21 (s,3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.90 (m, 2H), 1.83 (m, 2H), 1.45 (m,2H), 1.23 (m, 2H), 0.83 (t, 3H).

Step 9:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide(14 g, 0.023 mol) in dichloromethane (150 mL) was added aqueous 35%formaldehyde solution (2.4 g, 0.080 mol) at 0° C. After stirring for 20min, Na(OAc)₃BH (12.2 g, 0.057 mol) was added and stirring continued for2 h at 0° C. Water (100 mL) was then added to the reaction mixture andthe mixture extracted with 20% methanol in DCM (3×200 mL). The combinedextracts were dried over Na₂SO₄ and the solvent removed under reducedpressure. The crude product was purified by basic alumina columnchromatography eluting with 6-7% MeOH in DCM to afford the titlecompound (10 g, 63.6%). LCMS: 614.65 (M+1)+; HPLC: 98.88% (@ 210-370 nm)(R_(t); 3.724; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.45 (s, 1H), 8.17 (t, 1H), 7.56 (d, 2H, J=8 Hz), 7.36 (m, 3H), 7.17(s, 1H), 5.85 (s, 1H), 4.29 (d, 2H, J=4.4 Hz), 3.57 (bs, 4H), 3.48 (s,2H), 3.09 (q, 2H), 2.66 (m, 1H), 2.36 (bs, 4H), 2.21 (s, 3H), 2.20 (s,3H), 2.11 (s, 9H), 1.79 (m, 4H), 1.36 (m, 2H), 1.11 (m, 2H), 0.82 (t,3H, J=6.4&6.8 Hz).

Example 142: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-((2-methoxyethyl)(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-((2-methoxyethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (400 mg,1.22 mmol) and 2-methoxyacetaldehyde (1.3 mg, 17.56 mmol) in 7 ml ofdichloroethane, acetic acid (0.42 mL, 7.33 mmol) was added and stirredat room temperature for 20 minutes. The reaction mixture was cooled to0° C. and sodium triacetoxyborohydride (777 mg, 3.66 mmol) was added andstirred at room temperature for 2 h. The reaction mixture was thenneutralized with sat. NaHCO₃ and extracted with DCM, and the organiclayer was dried over Na₂SO₄, concentrated under reduced pressure to get260 mg of crude product.

Step 2: Synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((2-methoxyethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

A mixture of methyl5-bromo-3-((2-methoxyethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate(260 mg, 0.67 mmol) and NaOH (40 mg, 1.01 mmol) in 5 ml of ethanol:water(2:1) was heated at 70° C. for 2 h. The reaction mixture wasconcentrated to dryness and the crude dissolved in water, pH wasadjusted to 5 to 6 by slow addition of HCl and compound was extracted in10% MeOH in DCM. Organic layer was dried over Na₂SO₄, concentrated underreduced pressure to afford 230 mg of acid.

The crude acid (230 mg, 0.62 mmol),3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (188 mg, 1.24 mmol), PyBOP(483 mg, 0.93 mmol) and triethyl amine (0.17 ml, 1.238) mixture wasstirred in 3 ml of DMSO at rt for overnight. The reaction mixture wasdiluted with water and compound was extracted in 10% MeOH in DCM, driedover Na₂SO₄, concentrated and crude was purified by silica gel (100-200)column chromatography to get5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((2-methoxyethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(110 mg, 35%).

Step 3: SynthesisN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-((2-methoxyethyl)(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((2-methoxyethyl)(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(110 mg, 0.21 mmol), (4-(morpholinomethyl)phenyl)boronic acid (99 mg,0.33 mmol), sodium carbonate (83 mg, 0.78 mmol) in 4 ml of dioxane wasdegassed with argon for 20 min, Pd(PPh₃) (25 mg, 0.02 mmol) was added tothe mixture and heated to 100° C. for overnight. The reaction was cooledto room temperature and diluted with water, compound was extracted in10% MeOH in DCM, dried over Na₂SO₄, concentrated and crude productpurified by silica gel (100-200) chromatography to obtain the titlecompound (50 mg 38%).

LCMS: 603.45 (M+1)⁺; HPLC: 99.60% (@ 254 nm) (R_(t); 4.492; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (bs, 1H), 8.20(t, 1H), 7.58 (d, 2H, J=7.2 Hz), 7.47 (s, 1H), 7.37 (d, 2H, J=7.2 Hz),7.23 (s, 1H), 5.86 (s, 1H), 4.29 (d, 2H, J=3.6 Hz), 3.82-3.85 (m, 2H),3.49-3.58 (m, 6H), 3.15-3.3.23 (m, 9H), 2.98 (m, 1H), 2.36 (m, 4H), 2.23(s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.51-1.68 (m, 4H).

Example 143:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl-d5(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl 5-bromo-3-(ethyl-d5(tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzoate

To a stirred solution of compound methyl5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (Ig, 3.05mmol) and sodium triacetoxyborodeuteride (0.2 g, 4.76 mmol) indichloroethane (15 mL), acetic acid (1.65 g, 27.5 mmol) was added andreaction stirred at 5-10° C. for 2 h. Then acetaldehyde-d4 (0.264 g,6.00 mmol) was added at 0° C. and reaction stirred at room temperaturefor overnight. On completion, reaction was quenched with aqueous sodiumbicarbonate, organic phase was separated and aqueous phase was extractedwith dichloromethane. Combined organic layers were dried over sodiumsulphate and concentrated under reduced pressure to give crude materialwas purified by column chromatography to afford desired product (1 g,91%).

Steps 2 and 3: Synthesis of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl-d5 (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

Aqueous NaOH (0.166 g, 4.15 mmol) was added to a solution of compoundmethyl5-bromo-3-(ethyl-d5(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate (1g, 2.77 mmol) in ethanol (10 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand acidified using dilute HCl up to pH 6 and pH 4 was adjusted usingcitric acid. Extraction was carried out using ethyl acetate. Combinedorganic layers were dried concentrated giving respective acid (0.7 g,2.01 mmol, 73%), which was then dissolved in DMSO (7 mL) and 3-(aminomethyl)-4, 6-dimethylpyridin-2(1H)-one (0.611 g, 4.01 mmol) was added toit. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (1.56 g, 3.01 mmol) was added to it and stirring wascontinued for overnight. After completion of conversion, the reactionmass was poured into ice, extracted with 10% MeOH/DCM. Combined organiclayers were dried, concentrated to obtain crude; which then purified bysolvent washings to afford desired product (0.6 g, 62%).

Step 4: Synthesis of N-((4, 6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)methyl)-5-(ethyl-d5 (tetrahydro-2H-pyran-4-yl)amino)-4′-formyl-4-methyl-[1, 1′-biphenyl]-3-carboxamide

To a stirred solution of compound 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl-d5 (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (0.3 g, 0.62 mmol) and (4-(morpholinomethyl)phenyl) boronic acid (0.283 g, 0.93 mmol) in dioxane/water mixture,Na₂CO₃ (0.24 g, 2.26 mmol) was added and solution purged with argon for15 min. Then Pd (PPh₃)₄ (0.072 g, 0.062 mmol) was added and the mixturewas purged again for 10 min. Reaction mass was heated at 100° C. for 4h. On completion, reaction mixture was diluted with water and extractedwith 10% MeOH/DCM. Combined organic layers were dried over Na₂SO₄ andsolvent removed under reduced pressure to afford crude material whichwas purified by column chromatography over silica gel to afford desiredtitle compound (0.22 g, 61%). Analytical Data of N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl) methyl)-5-(ethyl-d5(tetrahydro-2H-pyran-4-yl) amino)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide: LCMS: 578.35 (M+1)⁺; HPLC: 98.50% (@ 254 nm)(R_(t); 4.176; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (bs, 1H), 8.19 (t, 1H), 7.57 (d, 2H, J=7.6), 7.36-7.39 (m, 3H),7.21 (s, 1H), 5.86 (s, 1H), 4.29 (d, 2H, J=3.2 Hz), 3.81-3.84 (m, 2H),3.48-3.57 (m, 6H), 3.22-3.25 (m, 2H), 3.02 (m, 1H), 2.36 (m, 4H), 2.24(s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.51-1.67 (m, 4H).

Example 144: Synthesis of5-((2,2-difluoroethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoate

In a 2-necked, 100 ml RBF, a stirred solution of methyl5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate (500 mg, 1.5 mmol) indifluoroacetic acid (15 ml), was treated with sodium tetrahydroborate(1000 mg, 26 mmol) portionwise over 12 minutes (CARE!). The reactionmixture was warmed to 50° C. and stirred for 4 hours. The reactionmixture was allowed to reach room temperature and then poured over ice(130 ml) and left for 5 minutes. The mixture was basified by theaddition of 6M NaOH (aq) (35 ml) and the pH adjusted to 7 using 1M HCl(aq) (20 ml). The resulting suspension was allowed to stand until thesolution was clear and the resulting solid collected by filtration anddried in-vacuo at 40° C. to give the title compound (572 mg, 96%) as awhite solid. LC-MS 100%, 2.32 min (3.5 minute LC-MS method), m/z=391.9,392.9, 393.9, 394.9; 1H NMR (500 MHz, Chloroform-d) δ ppm 7.79 (d,J=1.89 Hz, 1H) 7.44 (d, J=1.89 Hz, 1H) 5.44-5.71 (m, 1H) 4.00 (dd,J=11.51, 4.10 Hz, 2H) 3.91 (s, 3H) 3.41 (td, J=13.99, 4.18 Hz, 2H) 3.32(t, J=11.27 Hz, 2H) 2.97 (tt, J=11.37, 3.84 Hz, 1H) 2.47 (s, 3H)1.72-1.81 (m, 2H) 1.59-1.67 (m, 2H).

Step 2: Synthesis of5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoic acid

To a stirred solution of methyl5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoate (571mg, 1.5 mmol) in a mixture of THF (14.6 ml) and MeOH (2.2 ml), was added4M NaOH (14.6 ml). The reaction mixture was stirred at 50° C. for 7hours. The heat was switched off and the reaction mixture was stirred atroom temperature for 16.5 hours. THF was removed in-vacuo and theaqueous residue was acidified to pH 4 by the addition of 6M HCl (aq) (10ml) with ice cooling. The resulting solid was collected by filtrationand washed with water (20 ml), dried in-vacuo at 30-40° C. for 3 hoursto give the title compound (526 mg, 96%) as a light beige solid. LC-MS100%, 1.98 min (3.5 minute LC-MS method), m/z=377.9, 378.9, 379.9,380.9; ¹H NMR (500 MHz, Chloroform-d) δ ppm 7.91 (d, J=1.58 Hz, 1H) 7.49(d, J=1.58 Hz, 1H) 5.43-5.75 (m, 1H) 4.01 (dd, J=11.43, 3.55 Hz, 2H)3.42 (td, J=13.95, 3.78 Hz, 2H) 3.32 (t, J=11.35 Hz, 2H) 2.98 (tt,J=11.37, 3.53 Hz, 1H) 2.52 (s, 3H) 1.77 (d, J=10.88 Hz, 2H) 1.56-1.69(m, 2H). OH not visible.

Step 3: Synthesis of5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methylbenzamide

A stirred solution of5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoic acid(250 mg, 0.66 mmol) in dry DMF (3.0 ml) at 0° C. under a balloon ofnitrogen, was treated with HATU (327 mg, 0.86 mmol) and DIPEA (230 l,1.3 mmol) dropwise. The resulting solution was stirred for 5 minutes andthen treated with 3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one(89%, 136 mg, 0.79 mmol). The resulting suspension was stirred at 0° C.for 20 minutes and then stirred at room temperature overnight. After 18hours, 3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (25 mg) wasadded and stirring continued for a further 25 hours. The reactionmixture was diluted with water (30 ml) and CH₂Cl₂ (30 ml). The layerswere separated and the aqueous phase was extracted with CH₂Cl₂ (3×15ml). The combined organic phases were washed with a saturated solutionof NaHCO₃ (aq) (45 ml), water (2×50 ml), brine (2×50 ml), dried (MgSO₄),filtered and concentrated in-vacuo. The residue was purified by columnchromatography (10 g SNAP cartridge, Isolera, 0-3% MeOH:CH₂Cl₂) and thentriturated with ether. The resulting solid was collected by filtrationand dried in-vacuo at 40° C. to give the title compound (259 mg, 77%) asan off white solid. LC-MS 100%, 4.04 min (7 minute LC-MS method),m/z=512.0, 513.0, 514.0, 515.0; ¹H NMR (500 MHz, Acetone) δ 10.71 (s,1H), 7.57-7.49 (m, 2H), 7.25 (d, J=1.9 Hz, 1H), 5.91 (s, 1H), 5.76 (tt,J=56.2, 4.3 Hz, 1H), 4.40 (d, J=5.5 Hz, 2H), 3.88 (dd, J=11.3, 4.2 Hz,2H), 3.52 (td, J=14.6, 4.2 Hz, 2H), 3.33-3.23 (m, 2H), 3.02 (tt, J=11.6,3.9 Hz, 1H), 2.32 (s, 3H), 2.28 (s, 3H), 2.24 (s, 3H), 1.73 (dd, J=12.4,1.9 Hz, 2H), 1.59 (qd, J=12.2, 4.5 Hz, 2H).

Step 4: Synthesis of5-((2,2-difluoroethyl)(tetrahydro-2H-pyran-4-yl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

In a 2 necked RBF,5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methylbenzamide(200 mg, 0.39 mmol) and4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (130mg, 0.43 mmol) in dioxane (3.0 ml) were treated with a solution ofNa₂CO₃ (145 mg, 1.4 mmol) in water (1.0 ml). The mixture was brieflysonicated and nitrogen was bubbled through the resulting suspension witha long needle for 5 minutes. The suspension was then treated withpalladium-triphenylphosphane (1:4) (45 mg, 0.04 mmol) and nitrogen wasbubbled through the resulting suspension for a further 5 minutes. Thereaction mixture was then heated to 100° C. for 8 hours and stirred atroom temperature for 16 hours. The reaction mixture was treated withwater (20 ml) and 10% MeOH/CH₂Cl₂ (15 ml). The layers were separated andthe aqueous layer was extracted with 10% MeOH/CH₂Cl₂ (3×15 ml). Thecombined organics were washed with brine (55 ml), dried (MgSO₄),filtered and concentrated in-vacuo. The crude residue was purified bycolumn chromatography (10 g SNAP cartridge, Isolera, 0-5% MeOH/CH₂Cl₂)and treated with ether (10 ml), briefly sonicated, warmed in a waterbath and cooled on ice. The resulting white solid was collected byfiltration and washed with ether (5 ml). The solid was dried in-vacuo at40° C. for 35 hours to give the title compound (159 mg, 67%) as anoff-white solid. LC-MS 100%, 3.09 min (7 minute LC-MS method),m/z=609.15; ¹H NMR (500 MHz, Acetone) δ 10.72 (s, 1H), 7.63 (d, J=1.5Hz, 1H), 7.60 (d, J=8.1 Hz, 2H), 7.54 (t, J=5.2 Hz, 1H), 7.41 (d, J=1.6Hz, 1H), 7.39 (d, J=8.1 Hz, 2H), 5.91 (s, 1H), 5.76 (ttt, J=56.4, 30.4,4.4 Hz, 1H), 4.44 (d, J=5.5 Hz, 2H), 3.89 (dd, J=11.4, 3.9 Hz, 2H),3.65-3.55 (m, 6H), 3.50 (s, 2H), 3.29 (t, J=11.3 Hz, 2H), 3.07 (tt,J=11.5, 3.6 Hz, 1H), 2.40 (s, 4H), 2.37 (s, 3H), 2.34 (s, 3H), 2.22 (s,3H), 1.82-1.75 (m, 2H), 1.61 (qd, J=11.9, 4.1 Hz, 2H).

Example 145:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

Step 1: Synthesis of3-(((1s,4s)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

To a cooled solution of compound tert-butyl((1s,4s)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate (1.0 g, 1.60 mmol) in 10 ml of DCM, 2 ml of TFA was added dropwise and reaction mixture was stirred at rt for 2 h. Reaction mass wasconcentrated to dryness under reduced pressure, obtained crude wasdissolved in 10% MeOH in DCM and washed with sat NaHCO₃, water andbrine. Organic phase was dried over Na₂SO₄, concentrated under reducedpressure to obtain crude desired compound (650 mg, 81%).

Step 2: Synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide

To a stirred solution of crude compound3-(((1s,4s)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(650 mg, 1.32 mmol) and formaldehyde (0.5 ml of 38% solution, 13.26mmol) in 10 ml of methanol, sodium cyanoborohydride (82 mg, 1.32 mmol)was added at 0° C. and stirred at room temperature for overnight.Reaction mixture was partitioned between water and 10% MeOH in DCM,organic layer was dried over Na₂SO₄, concentrated under reducedpressure. Crude was purified by basic alumina column purification togive desired product compound (450 mg, 65%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzamide

A solution of compound5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(150 mg, 0.29 mmol), (1-methyl-1H-pyrazol-4-yl)boronic acid (72 mg, 0.34mmol), sodium carbonate (110 mg, 1.06 mmol) in 10 ml of dioxane wasdegassed with argon for 20 min, Pd(PPh₃) (33 mg, 0.03 mmol) was added tothe mixture and heated to 100° C. for overnight. Reaction was cooled toroom temperature and diluted with water, compound was extracted in 10%MeOH in DCM, dried over Na₂SO₄, concentrated and crude was purified bysilica gel (100-200) chromatography to obtain title compound (40 mg,26%).

Analytical Data: LCMS: 519.40 (M+1)⁺; HPLC: 95.98% (@ 254 nm) (R_(t);3.987; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A;0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col.Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min,Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45(bs, 1H), 8.12-8.09 (m, 2H), 7.80 (s, 1H), 7.29 (s, 1H), 7.10 (s, 1H),5.86 (s, 1H), 4.27 (d, 2H, J=4.8 Hz), 3.84 (s, 3H), 3.07-305 (m, 2H),2.67-2.63 (m, 2H), 2.21 (s, 3H), 2.15 (s, 3H), 2.12-2.11 (s, 3H+3H+3H),1.79-1.75 (m, 4H), 1.36-1.11 (m, 4H), 0.80 (t, 3H, J=6.0 Hz).

Example 146:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-((4-methyl-1,4-diazepan-1-yl)methyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide

A solution of compound5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(380 mg, 0.73 mmol), (4-formylphenyl)boronic acid (165 mg, 1.10 mmol),sodium carbonate (280 mg, 2.6 mmol) in 5 ml of dioxane was degassed withargon for 20 min, Pd(PPh₃) (84 mg, 0.07 mmol) was added to the mixtureand heated to 100° C. for 5 h. Reaction was cooled to room temperatureand diluted with water, compound was extracted in 10% MeOH in DCM, driedover Na₂SO₄, concentrated and crude was purified by silica gel (100-200)chromatography to obtain the desired compound (250 mg, 63%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-((4-methyl-1,4-diazepan-1-yl)methyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of compoundN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1s,4s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4′-formyl-4-methyl-[1,1′-biphenyl]-3-carboxamide(90 mg, 0.16 mmol) and 1-methyl-1,4-diazepane (0.56 g, 0.49 mmol) in 2ml of methanol, acetic acid (0.03 mL, 0.49 mmol) was added and stirredat room temperature for 20 minutes. Reaction mixture was cooled to 0° C.and sodium cyanoborohydride (25 mg, 0.41 mmol) was added and stirred atroom temperature for 4 h. Reaction mixture was neutralized with sat.NaHCO₃ and compound was extracted in DCM, dried over Na₂SO₄,concentrated under reduced pressure, purified by prep. HPLC to give thetitle compound (26 mg, 25%).

Analytical data of TFA salt: LCMS: 641.50 (M+1)⁺; HPLC: 97.72% (@ 254nm) (R_(t); 3.783; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-D6, 400 MHz)δ 11.45 (bs, 1H), 9.54 (s, 1H), 8.20 (s, 1H), 7.74 (d, J=7.6 Hz, 2H),7.57 (d, J=7.6 Hz, 2H), 7.42 (s, 1H), 7.26 (s, 1H), 5.87 (s, 1H), 4.29(d, J=4.0 Hz, 2H), 3.83 (m, 4H), 3.25 (m, 3H), 3.17-3.12 (m, 4H), 2.84(s, 3H), 2.69, 2.68 (2s, 6H), 2.24 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H),2.10-1.89 (m, 6H), 1.46-1.44 (m, 4H), 0.84 (t, J=7.2 Hz, 3H).

Example 147:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Step 1: methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-2-methylbenzoate

To a stirred solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)-cyclohexyl)amino)-2-methylbenzoate(10 g, 22.72 mmol) and acetaldehyde (2.99 g, 67.95 mmol) indichloroethane (100 mL), acetic acid (8.18 g, 136.33 mmol) was added.The reaction mixture was stirred at room temperature for 20 minutes.Then sodium triacetoxyborohydride (14.45 g, 68.16 mmol) was added at 0°C. and reaction mixture stirred overnight at room temperature. Thesolvent was removed under reduced pressure, the residue taken up inwater and extracted using 5% MeOH/DCM. The combined extracts were driedand concentrated to give methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-2-methylbenzoatewhich was used as is in further reactions (9 g, 84.66%).

Step 2: tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)amino)cyclohexyl)carbamate

Aqueous NaOH (1.15 g, 28.84 mmol) was added to a solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-2-methylbenzoate(9 g, 19.23 mmol) in ethanol (10 mL) and stirred at 60° C. for 1 h. Theethanol was removed under reduced pressure and acidified to pH 6 usingdilute HCl and then to pH 4 using citric acid. The mixture was extractedwith acetate. The combined organic layers were dried and concentrated togive the respective acid (8.6 g, 98.50%).

The above acid (8.6 g, 18.90 mmol) was dissolved in DMSO (7 mL) and3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (5.74 g, 37.80 mmol) wasadded. The reaction mixture was stirred at room temperature for 15 minbefore PyBOP (14.70 g, 28.35 mmol) was added. The reaction mixture wasstirred overnight. The reaction mixture was poured into ice water andextracted with 10% MeOH/DCM. The combined extracts were dried andconcentrated to obtain the crude product which purified by solventwashings to afford tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)amino)cyclohexyl)carbamate(10.2 g, 91.89%).

Step 3:3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methylphenyl)amino)cyclohexyl)carbamate(3 g, 5.10 mmol) was taken up in DCM (20 mL) to which TFA (5 mL) wasthen added. The reaction mixture was stirred at room temperature for 1h. The solvent was removed under reduced pressure and saturated NaHCO₃solution was added. The mixture was extracted with 10% MeOH/DCM and thecombined extracts washed with water and brine, dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(2.2 g, 87.50%).

Step 4:5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide

3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(2.2 g, 4.50 mmol) was dissolved in DCM (25 mL) and cooled to 0° C.;formalin (0.49 g, 16.26 mmol) was then added. The reaction mixture wasstirred at same temperature for 20 minutes. Sodium triacetoxyborohydride(2.39 g, 11.22 mmol) was then added and the reaction mixture stirred atroom temperature for 1 h. The solvent were removed under reducedpressure and water was added to the residue. The mixture was extractedusing 10% MeOH/DCM. The combined extracts were dried and concentratedunder reduced pressure giving5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(2.3 g, 98.71%) which was used as is in further reactions.

Step 5:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and phenylboronic acid (1.5 eq) in dioxane/water mixture, Na₂CO₃(3.6 eq) was added and the solution purged with argon for 15 min. Pd(PPh₃)₄ (0.1 eq) was then added and the reaction mixture again purgedwith argon for 10 min. The reaction mixture was heated at 100° C. for 2h. The reaction mixture was diluted with water and extracted with 10%MeOH/DCM. The combined extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by chromatography over silica gel to afford the title compoundas a TFA salt (0.07 g, 23.92%). LCMS: 515.45 (M+1)⁺; HPLC: 92.45% (@254nm) (R_(t); 4.672; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.48 (bs, 1H), 9.41 (bs, 1H), 8.23 (bs, 1H), 7.63 (d, 2H, J=4.8 Hz),7.50-7.20 (m, 5H), 5.86 (s, 1H), 4.29 (d, 2H), 3.12 (m, 3H), 2.68 (s,6H), 2.25 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.95 (m, 4H), 1.44 (m,4H), 0.84 (t, 3H).

Example 148:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4,4′-dimethyl-[1,1′-biphenyl]-3-carboxamide

Step 1:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4,4′-dimethyl-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and p-tolylboronic acid (1.5 eq) in dioxane/water mixture, Na₂CO₃(3.6 eq) was added and the solution purged with argon for 15 min. Pd(PPh₃)₄ (0.1 eq) was then added and the reaction mixture again purgedwith argon for 10 min. The reaction mixture was heated at 100° C. for 2h. The reaction mixture was diluted with water and extracted with 10%MeOH/DCM. The combined extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by chromatography over silica gel to afford the title compoundas a TFA salt (0.15 g, 51.30%). LCMS: 529.40 (M+1)⁺; HPLC: 93.61% (@254nm) (R_(t); 4.761; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹HNMR (DMSO-d₆, 400 MHz)δ 11.47 (bs, 1H), 9.40 (bs, 1H), 8.21 (bs, 1H), 7.53 (d, 2H, J=6.8 Hz),7.38 (bs, 1H), 7.26 (d, 2H, J=7.6 Hz), 5.86 (s, 1H), 4.29 (d, 2H, J=4Hz), 3.15 (m, 2H), 2.69 (s, 3H), 2.68 (s, 3H), 2.33 (s, 3H), 2.24 (s,3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.95 (m, 4H), 1.44 (m, 4H), 0.84 (t,3H).

Example 149:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-(trifluoromethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-(trifluoromethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and (4-(trifluoromethyl)phenyl)boronic acid (1.5 eq) indioxane/water mixture, Na₂CO₃ (3.6 eq) was added and the solution purgedwith argon for 15 min. Pd (PPh₃)₄ (0.1 eq) was then added and thereaction mixture again purged with argon for 10 min. The reactionmixture was heated at 100° C. for 2 h. The reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. The combined extracts weredried over Na₂SO₄ and the solvent removed under reduced pressure toafford the crude product which was purified by chromatography oversilica gel to afford the title compound as a TFA salt (0.08 g, 23.52%).LCMS: 583.45 (M+1)⁺; HPLC: 94.04% (@ 254 nm) (R_(t); 5.168; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.48 (s, 1H), 9.41(bs, 1H), 8.27 (bs, 1H), 7.88 (d, 2H, J=8 Hz), 7.81 (d, 2H, J=8 Hz),7.51 (s, 1H), 7.34 (s, 1H), 5.87 (s, 1H), 4.30 (d, 2H, J=4.4 Hz), 3.16(m, 3H), 2.85 (m, 1H), 2.69 (s, 3H), 2.68 (s, 3H), 2.26 (s, 3H), 2.21(s, 3H), 2.11 (s, 3H), 1.94 (m, 4H), 1.45 (m, 4H), 0.85 (t, 3H, J=6.8Hz).

Example 150:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-(methylsulfonyl)-[1,1′-biphenyl]-3-carboxamide

Step 1:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-(methylsulfonyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and (4-(methylsulfonyl)phenyl)boronic acid (1.5 eq) indioxane/water mixture, Na₂CO₃ (3.6 eq) was added and the solution purgedwith argon for 15 min. Pd (PPh₃)₄ (0.1 eq) was then added and thereaction mixture again purged with argon for 10 min. The reactionmixture was heated at 100° C. for 2 h. The reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. The combined extracts weredried over Na₂SO₄ and the solvent removed under reduced pressure toafford the crude product which was purified by chromatography oversilica gel to afford the title compound as a TFA salt (0.12 g, 34.68%).LCMS: 593.45 (M+1)⁺; HPLC: 98.74% (@ 254 nm) (R_(t); 4.194; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.48 (bs, 1H), 9.43(s, 1H), 8.26 (s, 1H), 7.99 (d, 2H, J=8.4 Hz), 7.93 (d, 2H, J=8.4 Hz),7.51 (s, 1H), 7.34 (s, 1H), 5.87 (s, 1H), 4.30 (d, 2H, J=4.4 Hz), 3.24(s, 3H), 3.30 (m, 3H), 2.80 (m, 1H), 2.69 (s, 3H), 2.68 (s, 3H), 2.26(s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.93 (m, 4H), 1.45 (m, 4H), 0.84(t, 3H, J=6.8 Hz).

Example 151:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(pyrimidin-5-yl)benzamide

Step 1:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(pyrimidin-5-yl)benzamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and pyrimidin-5-ylboronic acid (1.5 eq) in dioxane/water mixture,Na₂CO₃ (3.6 eq) was added and the solution purged with argon for 15 min.Pd (PPh₃)₄ (0.1 eq) was then added and the reaction mixture again purgedwith argon for 10 min. The reaction mixture was heated at 100° C. for 2h. The reaction mixture was diluted with water and extracted with 10%MeOH/DCM. The combined extracts were dried over Na₂SO₄ and the solventremoved under reduced pressure to afford the crude product which waspurified by chromatography over silica gel to affordN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(pyrimidin-5-yl)benzamideTFA salt (0.12 g, 39.33%). LCMS: 517.60 (M+1)⁺; HPLC: 99.55% (@ 210nm-370 nm) (R_(t); 3.996; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% Bto 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.47 (bs, 1H), 9.43 (bs, 1H), 9.18 (s, 1H), 9.14 (s, 2H),8.22 (s, 1H), 7.59 (s, 1H), 7.41 (s, 1H), 5.87 (s, 1H), 4.30 (d, 2H),3.14 (m, 3H), 2.69 (s, 3H+3H), 2.27 (s, 3H), 2.22 (s, 3H), 2.11-2.07 (m,4H), 1.95 (m, 4H), 1.44 (m, 4H), 0.84 (t, 3H).

Example 152: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-5-(furan-2-yl)-2-methylbenzamideTFA salt

Step 1: synthesis of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)amino)-2-methylbenzoate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (5.0 g,2.0 mmol) and 4-N-Boc-aminocyclohexanone (5.69 g, 2.67 mmol) indichloroethane (50 mL) was added acetic acid (7.4 g, 12 mmol). Sodiumtriacetoxyborohydride (13.1 g, 6.17 mmol) was added at 0° C. and themixture was stirred at room temperature for 16 hours. The reactionmixture was quenched with aqueous sodium bicarbonate, the organic phaseseparated and the aqueous phase extracted with dichloromethane. Thecombined organic layers were dried over anhydrous sodium sulphate andconcentrated in-vacuo. The crude compound was purified by columnchromatography over silica gel (100-200 mesh size) eluting with 10%ethyl acetate in hexane to afford 3.5 g of the more polar trans-isomer5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)amino)-2-methylbenzoate(38%) of isomer as an off-white solid. ¹H NMR (CDCl₃, 400 MHz) δ 7.21(s, 1H), 6.89 (s, 1H), 4.41 (bs, 1H), 3.85 (s, 3H), 3.41-3.64 (m, 2H),2.11-2.21 (m, 6H), 1.42 (s, 9H), 1.22-1.36 (m, 5H),

Step 2: synthesis of synthesis of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)(ethyl)-amino)-2-methylbenzoate

To a stirred solution of5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)amino)-2-methylbenzoate(55 g, 0.12 mol) and acetaldehyde (11 g, 0.25 mol) in dichloroethane(550 mL) was added acetic acid (44.6 g, 0.74 mol). Sodiumtriacetoxyborohydride (79 g, 0.37 mol) was added at 0° C. and themixture was stirred at room temperature for 16 hours. Aqueous sodiumbicarbonate was added, the organic phase was separated and the aqueousphase extracted with dichloromethane. The combined organic layers weredried over anhydrous sodium sulphate and concentrated in-vacuo. Thecrude compound was purified by column chromatography over silica get toafford the title compound (35 g, 59%) as an off-white solid.

Step 3: synthesis of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)carbamoyl)-2-methylphenyl(ethyl)amino)cyclohexyl)carbamate

To a stirred solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)(ethyl)-amino)-2-methylbenzoate(25 g, 0.053 mol) in EtOH (100 mL) was added aqueous NaOH (3.5 g, 0.08mol in 10 mL H₂O). After stirring at 60° C. for 1 h, the mixture wasacidified to pH 4 and extracted with 10% methanol in DCM. The combinedorganic layers were dried and concentrated to give 24.2 g of thecorresponding acid. To a stirred solution of the acid (24 g, 0.053 mol)and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (16 g, 0.11 mol) andtriethyl amine (5.3 g, 0.053 mmol) in DMSO (50 mL) was added PyBop (41g, 0.079 mol). After stirring overnight at room temperature, the mixturewas poured into ice water and extracted with 10% MeOH/DCM. The combinedorganic layers were dried over sodium sulphate and concentrated underreduced pressure. The crude material was washed water (1 L×2) followedby acetonitrile (150 mL×3) to afford the title compound (24 g, 77%).

Step 4: synthesis of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

To a stirred solution of tert-butyl((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)carbamoyl)-2-methylphenyl(ethyl)amino)cyclohexyl)carbamate(6.0 g, 10 mol) in DCM (30 mL), was added TFA (10 mL) at 0° C. Thereaction mixture was stirred for 2 h at room temperature andconcentrated to dryness. The residue was neutralized by addition ofsaturated bicarbonate solution (40 mL) followed by extraction with 20%methanol in DCM (100 mL×4). The combined organic phases were dried overNa₂SO₄ and the solvent was removed under reduced pressure to afford 5.0g of the title compound which was used without further purification.

Step 4: synthesis of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide

To a stirred solution of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(5.0 g, 10 mmol) in dichloromethane (50 mL) was added aq. 35%formaldehyde solution (2.9 g, 36 mmol) at 0° C. Na(OAc)₃BH (5.43 g, 25.6mmol) was added and the mixture was stirred for 2 h at 0° C. Water (100mL) was added followed by extraction with 20% methanol in DCM (200mL×3). The combined organic layers were dried over Na₂SO₄ and thesolvent was removed under reduced pressure. The residue was purified bycolumn chromatography over basic alumina eluting with 6-7% MeOH in DCMto afford the title compound (4.5 g, 94%).

General Suzuki reaction procedure for the synthesis of Compounds152-156, 158-162, 165, and 167

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide (1 eq.)and given boronic acids/pinacol esters (1.2 eq.) in dioxane/watermixture (4:1), was added Na₂CO₃ (3.6 eq.). The solution was purged withargon for 15 min. Pd(PPh₃)₄ (0.1 eq.) was added. The stirred reactionmixture was heated at 100° C. for 2-4 h under argon. After cooling toroom temperature, the mixture was diluted with 10% MeOH/DCM andfiltered. The filtrate was concentrated, diluted with water andextracted with 10% MeOH in DCM. The combined organic layers were driedover Na₂SO₄ and the solvent removed under reduced pressure. The crudeproducts were purified either by column chromatography over silica gel(100-200 mesh) or by preparative HPLC to give products as free base orTFA salt respectively.

Analytical Data ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-5-(furan-2-yl)-2-methylbenzamideTFA salt (0.08 g, 27%); LCMS: 505.55 (M+1)⁺; HPLC: 97.76% (@ 210 nm-370nm) (R_(t); 4.192; Method: Column: YMC ODS-A 150 mm×4.6 mm×5 L; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.46 (bs, 1H), 8.21 (s, 1H), 8.17 (s, 1H), 7.71 (s, 1H), 7.42 (s,1H), 7.25 (s, 1H), 6.93 (s, 1H), 6.57 (s, 1H), 5.86 (s, 1H), 4.29 (d,2H), 3.08-3.06 (m, 3H), 2.67 (m, 1H), 2.21 (s, 3H+3H+3H), 2.18-2.11 (s,3H+3H), 1.80 (m, 4H), 1.37-1.19 (m, 4H), 0.81 (t, 3H).

Example 153:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(quinolin-8-yl)-benzamideTFA salt (0.09 g, 27%)

Analytical Data of TFA salt: LCMS: 566.70 (M+1)⁺; HPLC: 93.94% (@ 210nm-370 nm) (R_(t); 4.352; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30 OC; Flow rate: 1.4 mL/min.; Gradient: 5% B to95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.43 (bs, 1H), 9.34 (bs, 1H), 8.88 (s, 1H), 8.45 (d, 1H,J=7.6 Hz), 8.16 (t, 1H), 8.00 (d, 1H, J=7.2 Hz), 7.80-7.79 (m, 1H),7.70-7.69 (m, 1H), 7.59-7.57 (m, 1H), 7.49 (m, 1H), 7.29 (s, 1H), 5.85(s, 1H), 4.28 (d, 2H, J=3.6 Hz), 3.16-3.11 (m, 3H), 2.70-2.69 (m,1H+3H+3H), 2.30 (s, 3H), 2.19 (s, 3H), 2.09 (s, 3H), 1.99 (m, 4H), 1.45(m, 4H), 0.93 (t, 3H).

Example 154:5-(2-aminopyrimidin-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino-2-methylbenzamideTFA salt (0.14 g, 45%)

Analytical Data of TFA salt: LCMS: 532.65 (M+1)⁺; HPLC: 98.49% (@ 210nm-370 nm) (R_(t); 3.692; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30 OC; Flow rate: 1.4 mL/min.; Gradient: 5% B to95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.48 (bs, 1H), 9.45 (bs, 1H), 8.17 (s, 1H), 7.42 (s, 1H),7.24 (s, 1H), 5.87 (s, 1H), 4.28 (d, 2H, J=4.4 Hz), 3.12 (m, 3H), 2.69(s, 3H+3H), 2.23 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.96 (m, 4H), 1.43(m, 4H), 0.83 (t, 3H).

Example 155:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(pyridin-4-yl)-benzamideTFA salt (0.17 g, 56%)

Analytical Data of TFA salt: LCMS: 516.60 (M+1)⁺; HPLC: 92.58% (@ 210nm-370 nm) (R_(t); 3.775; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% Bto 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.45 (bs, 1H), 9.74 (bs, 1H), 8.85 (d, 2H, J=5.2 Hz), 8.30(t, 1H), 8.24 (d, 2H, J=4.8 Hz), 7.71 (s, 1H), 7.55 (s, 1H), 5.88 (s,1H), 4.31 (d, 2H, J=4.4 Hz), 3.16 (m, 3H), 2.79 (m, 1H), 2.69 (s,3H+3H), 2.28 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.98-1.90 (m, 4H),1.47-1.45 (m, 4H), 0.84 (t, 3H).

Example 156:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(thiophen-3-yl)-benzamideTFA salt (0.07 g, 56%)

Analytical Data of TFA salt: LCMS: 521.55 (M+1)⁺; HPLC: 98.64% (@ 210nm-370 nm) (R_(t); 4.366; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% Bto 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.47 (bs, 1H), 9.45 (bs, 1H), 8.18 (s, 1H), 7.87 (s, 1H),7.63 (s, 1H), 7.54 (m, 2H), 7.32 (s, 1H), 5.87 (s, 1H), 4.29 (d, 2H),3.13 (m, 3H), 2.69 (m, 6H+1H), 2.22 (s, 3H+3H), 2.11 (s, 3H), 1.96 (m,4H) 1.44 (m, 4H), 0.84 (t, 3H).

Example 158:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(6-methylpyridin-3-yl)benzamide

Analytical Data: LCMS: 530.55 (M+1)⁺; HPLC: 96.45% (@ 210 nm-370 nm)(R_(t); 4.192; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.45 (bs, 1H), 8.70 (s, 1H), 8.19 (s, 1H), 7.95-7.90 (d, 1H, J=8.0 Hz),7.39 (s, 1H), 7.35-7.30 (d, 1H, J=7.6 Hz), 7.22 (s, 1H), 5.86 (s, 1H),4.29 (d, 2H), 3.05-3.15 (m, 2H), 2.60-2.70 (m, 1H), Three protons mergedin solvent peak, 2.25-2.35 (m, 6H+1H), 2.0-2.25 (3H+3H+3H), 1.70-1.90(m, 4H), 1.30-1.20 (m, 2H), 1.0-1.20 (m, 2H), 0.75-0.85 (t, 3H)

Example 159:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-5-(3,5-dimethylisoxazol-4-yl)-2-methylbenzamideTFA salt (0.13 g, 50%)

Analytical Data of TFA salt: LCMS: 534.60 (M+1)⁺; HPLC: 96.65% (@ 210nm-370 nm) (R_(t); 4.352; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% Bto 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.46 (bs, 1H), 9.42 (s, 1H), 8.17 (s, 1H), 7.13 (s, 1H),6.92 (s, 1H), 5.85 (s, 1H), 4.25-4.30 (d, 2H), 3.0-3.20 (m, 3H),2.65-2.75 (m, 3H+3H), Three protons merged in solvent peak, 2.39 (s,3H), 2.05-2.25 (m, 3H+3H+3H+1H), 1.90-2.0 (m, 2H), 1.80-1.90 (m, 2H),1.35-1.50 (m, 4H), 0.80-0.90 (t, 3H).

Example 160:5-(1,5-dimethyl-1H-pyrazol-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamideTFA salt (0.06 g, 58%)

Analytical Data of TFA salt: LCMS: 533.80 (M+1)⁺; HPLC: 90.76% (@ 254nm) (R_(t); 5.583; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6,400 MHz)δ 11.45 (bs, 1H), 9.27 (s, 1H), 8.11 (s, 1H), 7.52 (s, 1H), 7.11 (s,1H), 6.95 (s, 1H), 5.85 (s, 1H), 4.20-4.30 (d, 2H), 3.76 (s, 2H),3.0-3.20 (m, 2H), 2.60-2.75 (m, 3H+3H), 2.33 (s, 3H), 2.19 (s, 3H), 2.10(s, 3H), 1.80-2.0 (m, 4H), 1.35-1.50 (m, 4H), 0.80-0.90 (t, 3H).

Example 161:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-5-(1-methylpyrazol-3-yl)-2-methylbenzamideTFA salt (0.1 g, 33%)

Analytical Data of TFA salt: LCMS: 519.45 (M+1)⁺; HPLC: 96.61% (@ 254nm) (R_(t); 6.026; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.47 (bs, 1H), 9.46 (s, 1H), 8.20 (s, 1H), 7.45 (s, 1H), 7.28 (s,1H), 7.06 (s, 1H), 6.38 (s, 1H), 5.86 (s, 1H), 4.20-4.25 (d, 2H), 3.83(s, 3H), 3.0-3.15 (m, 3H), 2.60-2.80 (m, 1H+3H+3H), 2.24 (s, 3H), 2.19(s, 3H), 2.10 (s, 3H), 1.80-2.0 (m, 2H+2H), 1.40-1.50 (m, 4H), 0.80-0.90(t, 3H).

Example 162:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-5-(pyridin-3-yl)-2-methylbenzamideTFA salt (0.1 g, 33%)

Analytical Data of TFA salt: LCMS: 516.50 (M+1)⁺; HPLC: 89.96% (@ 254nm) (R_(t); 6.026; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.46 (bs, 1H), 9.35 (s, 1H), 8.96 (s, 1H), 8.63 (s, 1H), 8.22 (m,2H), 7.61 (s, 1H), 7.51 (s, 1H), 7.33 (s, 1H), 5.86 (s, 1H), 4.25-4.35(d, 2H), 3.05-3.15 (m, 3H), 2.6-2.80 (m, 1H+3H+3H), 2.25 (s, 3H), 2.21(s, 3H), 2.10 (s, 3H), 1.90-2.0 (m, 2H+2H), 1.40-1.50 (m, 4H), 0.80-0.90(t, 3H).

Example 163: NN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-4′-(2H-tetrazol-5-yl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2H-tetrazole(1.5 eq) in dioxane/water mixture, Na₂CO₃ (3.6 eq) was added and thesolution purged with argon for 15 min. Pd (PPh₃)₄ (0.1 eq) was thenadded and the reaction mixture again purged with argon for 10 min. Thereaction mixture was heated at 100° C. for 2 h. The reaction mixture wasdiluted with water and extracted with 10% MeOH/DCM. The combinedextracts were dried over Na₂SO₄ and the solvent removed under reducedpressure to afford the crude product which was purified bychromatography over silica gel then prep. HPLC to afford the titlecompound as a TFA salt (0.125 g, 35.50%). LCMS: 583.40 (M+1)⁺; HPLC:90.26% (@ 210-370 nm) (R_(t); 4.130; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (bs, 1H), 9.32 (bs, 1H),8.23-8.11 (m, 3H), 7.90 (d, 2H, J=7.2 Hz), 7.50 (s, 1H), 7.34 (s, 1H),5.87 (s, 1H), 4.30 (d, 2H, J=4.4 Hz), 3.59 (s, 1H), 3.13 (m, 3H),2.69-2.68 (m, 6H), 2.26-2.10 (m, 9H), 1.94 (m, 4H), 1.44 (m, 4H) 0.85(m, 3H).

Example 164:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(2-methylpyrimidin-5-yl)benzamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (1.5eq) in dioxane/water mixture, Na₂CO₃ (3.6 eq) was added and the solutionpurged with argon for 15 min. Pd (PPh₃)₄ (0.1 eq) was then added and thereaction mixture again purged with argon for 10 min. The reactionmixture was heated at 100° C. for 2 h. The reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. The combined extracts weredried over Na₂SO₄ and the solvent removed under reduced pressure toafford the crude product which was purified by chromatography oversilica gel then prep HPLC to afford the title compound as a TFA salt(0.08 g, 25.97%). LCMS: 531.65 (M+1)⁺; HPLC: 99.61% (@ 210-370 nm)(R_(t); 3.981; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.47 (bs, 1H),9.46 (bs, 1H), 9.00 (m, 2H), 8.20 (s, 1H), 7.53 (s, 1H),7.35 (s, 1H), 5.86 (s, 1H), 4.29 (m, 2H), 3.125-3.127 (m, 3H), 2.69-2.50(m, 10H), 2.25-2.10 (m, 9H), 1.94 (m, 4H), 1.43 (m, 4H), 0.83 (m, 3H).

Example 165:5-(1,3-dimethyl-1H-pyrazol-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino-2-methylbenzamideTFA salt (0.18 g, 69%)

Analytical Data of TFA salt: LCMS: 533.80 (M+1)⁺; HPLC: 87.18% (@210-370 nm) (R_(t); 3.946; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% Bto 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (D20-d6,400 MHz) δ 7.93 (s, 1H), 7.62-7.57 (m, 2H), 6.31 (s, 1H), 4.492-4.494(m, 2H), 3.92-3.80 (m, 6H), 3.33 (m, 1H), 2.82 (m, 6H), 2.39-2.28 (m,16H), 1.66 (m, 4H), 1.04 (m, 3H).

Example 166:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(thiazol-4-yl)benzamide

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide(1 eq) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (1.5eq) in dioxane/water mixture, Na₂CO₃ (3.6 eq) was added and the solutionpurged with argon for 15 min. Pd (PPh₃)₄ (0.1 eq) was then added and thereaction mixture again purged with argon for 10 min. The reactionmixture was heated at 100° C. for 2 h. The reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. The combined extracts weredried over Na₂SO₄ and the solvent removed under reduced pressure toafford the crude product which was purified by chromatography oversilica gel then prep HPLC to afford the title compound as a TFA salt(0.07 g, 28.40%). LCMS: 522.50 (M+1)⁺; HPLC: 99.22% (@ 210-370 nm)(R_(t); 4.114; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.48 (bs, 1H), 9.36 (bs, 1H), 9.18 (s, 1H), 8.19 (bs, 2H), 7.79 (s,1H), 7.58 (s, 1H), 5.87 (s, 1H), 4.30 (d, 2H, J=4.4 Hz), 3.11 (m, 3H),2.73-2.68 (m, 7H), 2.22 (s, 6H), 2.11 (s, 3H), 1.95 (m, 4H), 1.44 (m,4H), 0.83 (t, 3H).

Example 167:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(thiophen-2-yl)-benzamideTFA salt (0.05 g, 50%)

Analytical Data: LCMS: 521.55 (M+1)⁺; HPLC: 88.13% (@ 210-370 nm)(R_(t); 4.412; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.47 (bs, 1H), 9.32 (bs, 1H), 8.22 (t, 1H), 7.53-7.38 (m, 3H),7.20-7.13 (m, 2H), 5.87 (s, 1H), 4.28 (d, 2H, J=3.6 Hz), 3.10 (m, 3H),2.69-2.68 (m, 7H), 2.21 (s, 6H), 2.11 (s, 3H), 1.95-1.90 (m, 4H), 1.44(m, 4H), 0.83 (t, 3H).

Example 168: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(thiazol-2-yl)-benzamideTFA salt

Step 1: synthesis of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(thiazol-2-yl)benzoate

To a stirred solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate(0.5 g, 1.2 mmol) and 2-bromothiazole (0.22 g, 1.38 mmol) indioxane/water mixture, was added Cs₂CO₃ (0.94 g, 2.88 mmol) at roomtemperature. The solution was purged with argon for 15 min. andPdCl₂(PPh₃)₂ (0.08 g, 0.11 mmol) was added. The mixture was heated at100° C. for 3 h under argon, diluted with water and extracted with 10%MeOH/DCM. The combined organic layers were dried over Na₂SO₄ and thesolvent was removed under reduced pressure. The crude material waspurified by column chromatography over silica gel to afford the titlecompound (0.36 g, 71%).

Step 2: synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(thiazol-2-yl)-phenyl)-(ethyl)-amino)-cyclohexyl)-carbamate

To a stirred solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(thiazol-2-yl)benzoate(0.36 g, 0.76 mmol) in ethanol (5 mL) was added aqueous NaOH (0.064 g,1.60 mmol) at room temperature. The mixture was heated at 60° C. for 1h. and concentrated under reduced pressure. The concentrate wasacidified to pH 4 and extracted with ethyl acetate. The combined organiclayers were dried and concentrated to give 0.26 g of crude acid. To astirred solution of the crude acid (0.26 g, ca. 0.56 mmol) and 3-(aminomethyl)-4, 6-dimethylpyridin-2(1H)-one (0.17 g, 1.13 mmol) in DMSO (3mL) was added PYBOP (0.44 g, 0.85 mmol) at room temperature. Afterstirring overnight, the mixture was poured onto ice and extracted with10% MeOH/DCM. The combined organic layers were washed with water, dried,and concentrated under reduced pressure to give the title compound (0.15g) which was used directly in the next step.

Step 3: synthesis of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(thiazol-2-yl)benzamide

To a stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(thiazol-2-yl)-phenyl)-(ethyl)-amino)-cyclohexyl)-carbamate(0.15 g, 0.25 mmol) in dichloromethane (3 mL) was added TFA (1 mL). Themixture was stirred at room temperature for 1 h., concentrated underreduced pressure and NaHCO₃ solution was added to the concentrate. Afterextracting with 10% MeOH/DCM, the combined organic layers were washedwith water and brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to give 0.11 g of the title compound which was useddirectly for the next step.

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(thiazol-2-yl)-benzamideTFA salt

To a stirred solution of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(thiazol-2-yl)benzamide(0.1 g, 0.20 mmol) in methanol (3 mL) was added formalin (0.06 g, 2.0mmol) at 0° C. Sodium cyanoborohydride (0.025 g, 0.59 mmol) was added,and the mixture was stirred at room temperature for 1 h. The reactionmixture was quenched with water and extracted with 10% MeOH/DCM. Thecombined organic layers were dried and concentrated under reducedpressure to a solid which was purified by preparative HPLC to afford thetitle compound as a TFA salt (0.06 g, 56%). Analytical Data of TFA Salt:LCMS: 522.60 (M+1)⁺; HPLC: 92.00% (@ 210-370 nm) (R_(t); 4.255; Method:Column: YMC ODS-A 150 mm×4.6 mm×5 L; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (bs, 1H), 9.39(bs, 1H), 8.30 (t, 1H), 7.90 (d, 1H, J=3.2 Hz), 7.78 (d, 1H, J=2.4 Hz),7.71 (s, 1H), 7.47 (s, 1H), 5.88 (s, 1H), 4.30 (d, 2H, J=4 Hz), 3.11 (m,3H), 2.77-2.68 (m, 7H), 2.24 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H),1.96-1.89 (m, 4H), 1.45 (m, 4H), 0.84 (t, 3H, J=6.4 Hz).

Example 169: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)-benzamide

Step 1: synthesis of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)amino)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)benzoate

To a stirred solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate(0.5 g, 1.15 mmol) and 2-bromo-1-methyl-1H-imidazole (0.22 g, 1.38 mmol)in dioxane/water mixture was added Cs₂CO₃ (0.94 g, 2.88 mmol) underargon. PdCl₂ (PPh₃)₂ (0.08 g, 0.11 mmol) was added and the mixture washeated at 100° C. for 4 h under argon. Water was added and the mixturewas extracted with 10% MeOH/DCM. The combined organic layers were driedover Na₂SO₄ and the solvent was removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)amino)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)benzoate(0.22 g, 40%).

Step 2: synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)-phenyl)-(ethyl)-amino)cyclohexyl)carbamate

To a stirred solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)amino)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)benzoate(0.22 g, 0.47 mmol) in ethanol (3 mL) was added aqueous NaOH (0.028 g,0.70 mmol). After stirring at 60° C. for 1 h., the mixture wasconcentrated under reduced pressure, acidified to pH 4 and extractedwith ethyl acetate. The combined organic layers were dried andconcentrated to give 0.16 g of crude acid. To a stirred solution of thecrude acid (0.16 g, 0.35 mmol) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.11 g, 0.70 mmol) in DMSO (3 mL) was addedPYBOP (0.27 g, 0.53 mmol). After stirring overnight the mixture waspoured into ice water and extracted with 10% MeOH/DCM. The combinedorganic layers were washed with water, dried and concentrated underreduced pressure to afford 0.12 g of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)-phenyl)-(ethyl)-amino)cyclohexyl)carbamatewhich was used directly without further purification.

Synthesis of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)benzamide

To a stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)-phenyl)-(ethyl)-amino)cyclohexyl)carbamate(0.12 g, 0.20 mmol) in DCM (3 mL) was added TFA (1 mL). After stirringfor 1 h. at room temperature, the mixture was concentrated under reducedpressure. Saturated NaHCO₃ solution was added to the residue followed byextraction with 10% MeOH/DCM. The combined organic layers were washedwith water and brine; dried and concentrated under reduced pressure togive 0.1 g of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)benzamidewhich was used directly without further purification.

Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)benzamideTFA salt

To a stirred solution of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(1-methyl-1H-imidazol-2-yl)benzamide(0.1 g, 0.20 mmol) in methanol (3 mL) was added formalin (0.06 g, 2.0mmol) at 0° C. Sodium cyanoborohydride (0.025 g, 0.59 mmol) was added,and the mixture was stirred at room temperature for 1 h. Water was addedfollowed by extraction with 10% MeOH/DCM. The combined organic layerswere dried and concentrated under reduced pressure to a solid which waspurified by preparative HPLC to afford the title compound as a TFA salt(0.03 g, 28%). Analytical Data of TFA Salt: LCMS: 519.65 (M+1)⁺; HPLC:96.10% (@ 254 nm) (R_(t); 3.976; Method: Column: YMC ODS-A 150 mm×4.6mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient:5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.47 (bs, 1H), 9.60 (bs, 1H), 8.24 (bs, 1H), 7.82(s, 1H), 7.78 (s, 1H), 7.52 (s, 1H), 7.35 (s, 1H), 5.87 (s, 1H), 4.30(bs, 2H), 3.86 (s, 3H), 3.10 (m, 3H), 2.69 (bs, 7H), 2.28 (s, 3H), 2.22(s, 3H), 2.11 (s, 3H), 1.96-1.88 (m, 4H), 1.46 (m, 4H), 0.84 (bs, 3H).

Example 170:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)benzamide

Compound 170 was prepared with the method similar to that described inExample 183 below.

Analytical Data: LCMS: 614.75 (M+1)⁺; HPLC: 98.17% (@ 210-370 nm)(R_(t); 3.598; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (bs, 1H), 10.10 (bs, 1H), 9.68 (bs, 1H), 8.21 (m, 2H), 7.47 (s,1H), 7.38 (s, 1H), 7.16 (s, 1H), 7.05 (d, 1H, J=4.8 Hz), 5.87 (s, 1H),4.54-4.51 (m, 2H), 4.30 (d, 2H, J=4 Hz), 3.53 (m, 2H), 3.13 (m, 7H),2.86 (s, 3H), 2.76-2.68 (m, 7H), 2.24 (s, 3H), 2.22 (s, 3H), 2.11 (s,3H), 1.97-1.90 (m, 4H), 1.43 (m, 4H), 0.83 (t, 3H, J=6.4 Hz).

Example 171:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)benzamide

Compound 171 was prepared with the method similar to that described inExample 183 below.

Analytical Data: LCMS: 573.75 (M+1)⁺; HPLC: 95.92% (@ 210-370 nm)(R_(t); 3.891; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (bs, 1H), 8.19 (t, 1H), 8.14 (d, 1H, J=4.8 Hz), 7.44 (s, 1H), 7.32(s, 1H), 6.97 (s, 1H), 6.88 (d, 1H, J=4.8 Hz), 5.86 (s, 1H), 4.29 (d,2H, J=4.4 Hz), 3.84-3.81 (m, 2H), 3.54 (m, 4H), 3.28-3.22 (m, 2H),3.10-3.02 (m, 3H), 2.42 (m, 4H), 2.24 (s, 3H), 2.23 (s, 3H), 2.10 (s,3H), 2.10 (s, 3H), 1.66-1.50 (m, 4H), 0.82 (t, 3H, J=6.4 Hz).

Example 172:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide

Compound 172 was prepared with the method similar to that described inExample 183 below.

Analytical Data: LCMS: 600.75 (M+1)⁺; HPLC: 99.58% (@ 210-370 nm)(R_(t); 3.460; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (bs, 1H), 9.59 (bs, 1H), 8.92 (bs, 2H), 8.47 (s, 1H), 8.16 (s,1H), 7.92 (d, 1H, J=7.6 Hz), 7.39 (bs, 1H), 7.21 (bs, 1H), 7.01 (d, 1H,J=8.8 Hz), 5.87 (s, 1H), 4.29 (d, 2H, J=5.2 Hz), 3.75 (q, 4H, J=5.2 Hz),3.22 (m, 4H), 3.12 (m, 3H), 2.75 (m, 1H), 2.69 (s, 3H), 2.68 (s, 3H),2.23 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.97 (m, 4H), 1.44 (m, 4H),0.83 (t, 3H, J=6.8 Hz).

Example 173:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide

Compound 173 was prepared with the method similar to that described inExample 183 below.

Analytical Data: LCMS: 559.55 (M+1)⁺; HPLC: 98.43% (@ 210-370 nm)(R_(t); 3.731; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.45 (bs, 1H), 8.86 (s, 2H), 8.48 (bs, 1H), 8.21 (bs, 1H), 7.94 (bs,1H), 7.45 (bs, 1H), 7.25 (bs, 1H), 7.02 (d, 1H, J=8.4 Hz), 5.87 (s, 1H),4.29 (d, 2H, J=3.6 Hz), 3.83 (m, 3H), 3.76 (bs, 4H), 3.30-3.15 (m, 7H),3.10 (m, 1H), 2.25 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.75-1.50 (m,4H), 0.84 (t, 3H).

Example 174: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(pyrazin-2-yl)-benzamide

Step 1: synthesis of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)(ethyl)-amino)-2-methylbenzoate

To a stirred solution of5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)amino)-2-methylbenzoate(10 g, 23 mmol) and acetaldehyde (2.99 g, 68 mmol) in dichloroethane(100 mL), was added acetic acid (8.18 g, 136 mmol) and reaction stirredat room temperature for 20 minutes. Sodium triacetoxyborohydride (14.45g, 68 mmol) was added at 0° C. and the mixture was stirred overnight atroom temperature. The solvent was removed under reduced pressure andwater was added followed by extraction with 5% MeOH/DCM. The combinedorganic layers were dried and concentrated to give 9 g of the titlecompound which was used without further purification.

Step 2: synthesis of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate

To a stirred solution of methyl5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)(ethyl)-amino)-2-methylbenzoate(2.0 g, 4.3 mmol) and bis pinacolatodiboron (5.42 g, 21 mmol) in dioxanewas added potassium acetate (1.25 g, 12.82 mmol) under argon.PdCl₂(dppf)DCM (0.35 g, 0.42 mmol) was added and the mixture was heatedat 80° C. for 3 h under argon. Water was added followed by extractionwith ethyl acetate. The combined organic layers were dried over Na₂SO₄,and the solvent was removed under reduced pressure. The crude productwas purified by column chromatography over silica gel to afford thetitle compound (1.3 g, 70%).

Step 3: synthesis of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(pyrazin-2-yl)benzoate

To a stirred solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate(0.50 g, 1.15 mmol) and 2-bromopyrazine (0.24 g, 1.49 mmol) indioxane/water mixture) was added CS₂CO₃ (0.94 g, 2.89 mmol under argon.PdCl₂ (PPh₃)₂ (0.08 g, 0.11 mmol) was added and the mixture was heatedat 100° C. for 3 h. under argon. Water was added followed by extractionwith 10% MeOH/DCM. The combined organic layers were dried over Na₂SO₄and the solvent removed under reduced pressure. The crude product waspurified by column chromatography over silica gel to afford the titlecompound (0.29 g, 53%).

Step 4: synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)carbamoyl)-2-methyl-5-(pyrazin-2-yl)-phenyl)-(ethyl)-amino)-cyclohexyl)-carbamate

To a stirred solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(pyrazin-2-yl)benzoate(0.29 g, 0.62 mmol) in ethanol (3 mL) was added aqueous NaOH (0.037 g,0.93 mmol) at room temperature, After stirring at 60° C. for 1 h., themixture was concentrated under reduced pressure, acidified to pH 4 andextracted with ethyl acetate. The combined organic layers were dried andconcentrated to give 0.24 g of crude acid. To a stirred solution of thecrude acid (0.24 g, 0.52 mmol) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.16 g, 1.05 mmol) in DMSO (3 mL) was addedPYBOP (0.41 g, 0.79 mmol). After stirring overnight the mixture waspoured into ice water and extracted with 10% MeOH/DCM. The combinedorganic layers were washed with water, dried and concentrated underreduced pressure to give 0.3 g of the title compound which was useddirectly without further purification.

Step 5: synthesis of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(pyrazin-2-yl)benzamide

To a stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)carbamoyl)-2-methyl-5-(pyrazin-2-yl)-phenyl)-(ethyl)-amino)-cyclohexyl)-carbamate(0.3 g, 0.51 mmol) in DCM (3 mL) was added TFA (1 mL). After stirring atroom temperature for 1 h., the mixture was concentrated under reducedpressure. Saturated NaHCO₃ solution was added followed by extractionwith 10% MeOH/DCM. The combined organic layers were washed with waterand brine, dried and concentrated under reduced pressure to give 0.24 gof the title compound which used directly without further purification.

Step 6: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(pyrazin-2-yl)benzamideTFA salt

To a stirred solution of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(pyrazin-2-yl)benzamide(0.24 g, 0.51 mmol) in methanol (3 mL) was added formalin (0.15 g, 5.1mmol) at 0° C. Sodium cyanoborohydride (0.06 g, 1.0 mmol) was added, andthe mixture was stirred at room temperature for 1 h. Water was addedfollowed by extraction with 10% MeOH/DCM. The combined organic layerswere dried and concentrated under reduced pressure. The solid obtainedwas purified by preparative HPLC to afford the title compound as a TFAsalt (0.12 g, 47%). Analytical Data of TFA Salt: LCMS: 517.50 (M+1)⁺;HPLC: 99.49% (@ 210-370 nm) (R_(t); 4.072; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.47 (bs, 1H), 9.45 (bs, 1H), 9.26(s, 1H), 8.71 (s, 1H), 8.61 (s, 1H), 8.23 (t, 1H), 7.92 (s, 1H), 7.74(s, 1H), 5.88 (s, 1H), 4.31 (d, 2H, J=4 Hz), 3.13 (m, 3H), 2.78 (m, 1H),2.69 (d, 6H, J=4.8 Hz), 2.28 (s, 3H), 2.22 (s, 3H), 2.12 (s, 3H),1.96-1.92 (m, 4H), 1.45 (m, 3H), 0.84 (t, 3H, J=6.4 Hz).

Example 175: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(5-methylpyrazin-2-yl)-benzamide

Step 1: synthesis of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(5-methylpyrazin-2-yl)benzoate

To a stirred solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate

(0.40 g, 0.92 mmol) and 2-bromo-5-methylpyrazine (0.21 g, 1.19 mmol) indioxane/water mixture was added Cs₂CO₃ (0.75 g, 2.30 mmol) under argon.PdCl₂ (PPh₃)₂ (0.064 g, 0.092 mmol) was added and the mixture was heatedat 100° C. for 3 h. under argon. Water was added followed by extractionwith 10% MeOH/DCM. The combined organic layers were dried over Na₂SO₄and the solvent was removed under reduced pressure to afford crudematerial which was purified by column chromatography over silica gel toafford the title compound (0.3 g, 56%).

Step 2: synthesis of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(5-methylpyrazin-2-yl)-phenyl)-(ethyl)-amino)-cyclohexyl)carbamate

To a solution of methyl3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-2-methyl-5-(5-methylpyrazin-2-yl)benzoate(0.29 g, 0.49 mmol) in ethanol (3 mL)) was added aqueous NaOH (0.029 g,0.75 mmol) at room temperature. After stirring at 60° C. for 1 h., themixture was concentrated under reduced pressure, acidified to pH 4 andextracted with ethyl acetate. The combined organic layers were dried andconcentrated to give 0.25 g of crude acid. To a stirred solution of thecrude acid (0.25 g, 0.44 mmol) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.13 g, 0.88 mmol) in DMSO (3 mL) was addedPYBOP (0.34 g, 0.66 mmol) at room temperature. After stirring wascontinued for overnight, the mixture was poured into ice water andextracted with 10% MeOH/DCM. The combined organic layers were washedwith water, dried and concentrated under reduced pressure to give 0.2 gof the title compound which was used directly without furtherpurification.

Step 3: synthesis of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(5-methylpyrazin-2-yl)benzamide

To a stirred solution of tert-butyl((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(5-methylpyrazin-2-yl)-phenyl)-(ethyl)-amino)-cyclohexyl)carbamate(0.2 g, 0.33 mmol) in DCM (3 mL) was added TFA (1 mL). After stirring atroom temperature for 1 h., the mixture was concentrated under reducedpressure. Saturated NaHCO₃ solution was added followed by extractionwith 10% MeOH/DCM. The combined organic layers were washed with waterand brine, dried and concentrated under reduced pressure to give 0.15 gof the title compound which was used directly without furtherpurification.

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(5-methylpyrazin-2-yl)-benzamideTFA salt

To a stirred solution of3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(5-methylpyrazin-2-yl)benzamide(0.15 g, 0.29 mmol) in methanol (3 mL) was added formalin (0.089 g, 2.98mmol) at 0° C. Sodium cyanoborohydride (0.037 g, 0.59 mmol) was addedand the mixture was stirred at room temperature for 1 h. Water was addedfollowed by extraction with 10% MeOH/DCM. The combined organic layerswere dried and concentrated under reduced pressure. The solid obtainedwas further purified by preparative HPLC to the title compound as a TFAsalt (0.12 g, 75%). Analytical Data of TFA Salt: LCMS: 531.50 (M+1)⁺;HPLC: 88.93% (@ 210-370 nm) (R_(t); 4.130; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.49 (bs, 1H), 9.54 (bs, 1H), 9.10(s, 1H), 8.59 (s, 1H), 8.23 (t, 1H), 7.88 (s, 1H), 7.69 (s, 1H), 5.87(s, 1H), 4.30 (bs, 2H), 3.12 (m, 3H), 2.77-2.69 (m, 7H), 2.53 (s, 3H),2.26 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.95-1.91 (m, 4H), 1.44 (m,4H), 0.83 (t, 3H).

Example 176: Synthesis of5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of6-methyl-2-oxo-4-propyl-1,2-dihydropyridine-3-carbonitrile

To a stirred solution of t-BuOK (Ig, 8.9 mmol) in DMSO (15 mL) at rt,was added compound cyanoacetamide (0.824 g, 9.8 mmol) and(E)-hept-3-en-2-one (Ig, 8.91 mmol). Reaction mixture was stirred for 30min at rt. Additional t-BuOK (3 g, 26.7 mmol) was added and reaction wasstirred at rt in presence of air. On completion, it was diluted with H₂Oand slowly by 4N HCl. Precipitated solid was filtered, washed with waterand dried. Crude product was triturated with ether to afford the titlecompound (0.5 g, 33%).

Step 2: Synthesis of 3-(aminomethyl)-6-methyl-4-propylpyridin-2(1H)-one

To a solution of6-methyl-2-oxo-4-propyl-1,2-dihydropyridine-3-carbonitrile (1.3 g, 7.38mmol) in methanol and aq. ammonia solution (50 mL, 9:1), catalyticamount of Raney Nickel was added. Reaction mass was stirred at roomtemperature under hydrogen pressure (balloon pressure) for 5 h. Oncompletion of reaction, it was filtered through celite bed and filtratewas concentrated under reduce pressure to afford the title compound (1.2g, 92%).

Step 3: Synthesis of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

Aqueous NaOH (2.36 g, 59.15 mmol) was added to a solution of methyl5-bromo-3-(ethyl (tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzoate (14g, 39.43 mmol) in ethanol (100 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand acidified using dilute HCl up to pH 6 and pH 4 was adjusted usingcitric acid. Extraction was carried out using ethyl acetate. Combinedorganic layers were dried concentrated giving respective acid (13.9 g,99%).

The above acid (0.6 g, 1.75 mmol) was then dissolved in DMSO (5 mL) and3-(aminomethyl)-6-methyl-4-propylpyridin-2(1H)-one (0.64 g, 3 mmol) andtriethyl amine (0.49 g, 5.26 mmol) was added to it. The reaction mixturewas stirred at room temperature for 15 min before PyBOP (1.36 g, 2.63mmol) was added to it and stirring was continued for overnight. Aftercompletion of the reaction, the reaction mixture was poured into ice,extracted with 10% MeOH/DCM. Combined organic layers were dried,concentrated to obtain crude; which then purified by solvent washings toafford 5 the title compound (0.75 g, 84.7%).

Step 4: Synthesis of5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

To a stirred solution of 5-bromo-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (0.3 g, 0.59 mmol) and 4-((morpholino)methyl)phenylboronicacid pinacol ester (0.22 g, 0.71 mmol) in dioxane/watermixture (5 mL+1 mL), Na₂CO₃ (0.23 g, 2.14 mmol) was added and solutionwas purged with argon for 15 min. Then Pd(PPh₃)₄ (0.068 g, 0.059 mmol)was added and argon was purged again for 10 min. Reaction mass washeated at 100° C. for 4 h. On completion, reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure to affordcrude material which was purified by column chromatography over silicagel to afford the title compound (0.25 g, 70%). LCMS: 601.55 (M+1)⁺;HPLC: 97.21% (@ 210-370 nm) (R_(t); 4.380; Method: Column: YMC ODS-A 150mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA inacetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (bs, 1H), 8.16 (bs, 1H), 7.57(d, 2H, J=7.6 Hz), 7.38 (t, 3H, J=6.8 Hz), 7.21 (s, 1H), 5.89 (s, 1H),4.30 (m, 2H), 3.84-3.82 (m, 2H), 3.57 (bs, 3H), 3.48 (s, 3H), 3.28-3.22(m, 2H), 3.09-3.02 (m, 3H), 2.36 (bs, 4H), 2.25 (s, 3H), 2.11 (s, 3H),1.67-1.54 (m, 6H), 0.93 (t, 3H, J=7 Hz), 0.84 (t, 3H). 2H merged insolvent peak.

Example 177: Synthesis ofN-((5-bromo-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of methyl5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxylate

To a stirred solution of methyl5-bromo-3-(ethyl-(tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzoate (1g, 2.82 mmol) and 4-((morpholino)methyl) phenylboronic acid pinacolester (1.03 g, 3.38 mmol) in dioxane/water mixture (10 mL+2 mL), Na₂CO₃(1.08 g, 10.14 mmol) was added and solution purged with argon for 15min. Then Pd (PPh₃)₄ (0.325 g, 0.28 mmol) was added and argon was purgedagain for 10 min. Reaction mass was heated at 100° C. for 2 h. Oncompletion, reaction mixture was diluted with water and extracted with10% MeOH/DCM. Combined organic layers were dried over Na₂SO₄ and solventremoved under reduced pressure to afford crude material which waspurified by column chromatography over silica gel to afford the titlecompound (0.75 g, 59%).

Step 2: Synthesis of5-bromo-4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile

To a stirred suspension of4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (5 g, 33.78 mmol)in AcOH (25 mL) was dropwise added bromine (2.5 mL) at rt. Resultingsolution was stirred for 1 h. Solvent was removed under reducedpressure. Obtained solid was recrystallised in hot EtOH and H₂O to givethe title compound as a white solid (5.5 g, 72%).

Step 3: Synthesis of3-(aminomethyl)-5-bromo-4,6-dimethylpyridin-2(1H)-one

To a stirred solution of5-bromo-4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (1 g, 4.44mmol) and NiCl2.6H₂O (0.21 g, 0.89 mmol) in methanol at 0° C., NaBH₄(0.68 g, 17.78 mmol) was added portion wise. Reaction mixture was thenstirred at rt for overnight period. On completion, it was acidifiedusing 3N HCl and stirred at rt for 3 h. Solvent was removed underreduced pressure. Residue was washed with diethyl ether and basifiedwith aq. NH₄OH. Compound was extracted in 10% MeOH in DCM and dried overanhydrous Na₂SO₄ to give the title compound (0.96 g, 94%) which was usedas such for coupling reaction.

Step 4: Synthesis ofN-((5-bromo-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Aqueous NaOH (0.06 g, 1.66 mmol) was added to a solution of compound 7(0.5 g, 1.11 mmol) in EtOH:H₂O (4:1) (10 mL) and stirred at 60° C. for 1h. After completion of the reaction, ethanol was removed under reducedpressure and reaction mass was acidified using dilute HCl up to pH 6 andpH 4 was adjusted using citric acid. Extraction was carried out using10% MeOH in DCM. Combined organic layers were dried and concentratedgiving respective acid (0.35 g, 72%).

The above acid (0.266 g, 0.61 mmol) was then dissolved in DMSO (2.5 mL)and 3-(aminomethyl)-5-bromo-4,6-dimethylpyridin-2(1H)-one (0.42 g, 1.83mmol) and triethyl amine (0.095 g, 0.91 mmol) were added to it. Thereaction mixture was stirred at room temperature for 15 min before PYBOP(0.63 g, 1.22 mmol) was added to it and stirring was continued forovernight. After completion of the reaction, reaction mass was pouredinto ice, extracted with 10% MeOH/DCM. Combined organic layers weredried, concentrated to obtain crude; which then purified by solventwashings to afford the title compound (0.035 g, 7.6%).

LCMS: 653.65 (M+1)⁺, HPLC: 89.23% (@ 210-370 nm) (R_(t); 4.421; Method:Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d6, 400 MHz) δ 9.88 (bs, 1H), 8.30(bs, 1H), 7.76 (m, 2H), 7.57 (d, 2H, J=7.6 Hz), 7.44 (bs, 1H), 7.27 (bs,1H), 4.39 (m, 4H), 3.99-3.96 (m, 5H), 3.84 (d, 2H, J=8.4 Hz), 3.65-3.62(m, 2H), 3.28-3.23 (m, 4H), 3.12 (m, 4H), 2.35 (s, 3H), 2.31 (s, 3H),2.26 (s, 3H), 1.65-1.55 (m, 4H), 0.84 (t, 3H).

Example 178:4-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Compound 178 was prepared with the method similar to that described inExample 177.

Analytical Data: LCMS: 593.60 (M+1)⁺; HPLC: 95.50% (@ 210-370 nm)(R_(t); 4.566; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (s, 1H), 8.34 (m, 1H), 7.61-7.24 (m, 6H), 5.86 (s, 1H), 4.29 (m,2H), 3.86-3.84 (m, 2H), 3.57-3.49 (m, 6H), 3.25-3.16 (m, 5H), 2.36 (m,4H), 2.21 (s, 3H), 2.10 (s, 3H), 1.68-1.58 (m, 4H), 0.86 (t, 3H).

Example 179: Synthesis of5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

Step 1: Synthesis of5-fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile

To a stirred solution of 2-cyanoacetamide (689 mg, 8.2 mmol) inanhydrous EtOH (7.0 ml) at 75° C., was added 3-fluoropentane-2,4-dione(880 mg, 7.5 mmol), followed by piperidine (96 μl, 0.97 mmol). Thereaction mixture was stirred at this temperature for 3 hours and thereaction mixture left to reach room temperature before being stored inthe refrigerator for 4 days. The beige solid was collected by filtrationand rinsed with cold EtOH (4×0.4 ml) until the filtrate ran clear. Theresulting beige solid was dried in-vacuo at 40° C. for 5 hours to givethe title compound (733 mg, 58%) as a beige solid. LC-MS 97%, 1.18 min(3.5 minute LC-MS method); m/z=166.9, ¹H NMR (500 MHz, Chloroform-d) δppm 13.67 (br. s., 1H) 2.46 (d, J=2.05 Hz, 3H) 2.45 (d, J=2.84 Hz, 3H).

Step 2: Synthesis of3-(aminomethyl)-5-fluoro-4,6-dimethyl-1,2-dihydropyridin-2-one

A solution of 0.05M5-fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (731 mg,4.4 mmol) in 1.75M NH₃/MeOH (87 ml) was passed through the H-Cube at 80°C. and 50 bar at a flow rate of 1 ml/min. The resulting solution wasconcentrated in-vacuo. The resulting solid was split into 2 batches and350 mg of the crude product was purified by column chromatography (25 gSNAP cartridge, Isolera, 0-25% MeOH (containing 10% NH₄OH):CH₂Cl₂) togive the title compound (307 mg, 20%) as an off white solid and a 1:1mixture of product:starting material. LC-MS (ELS) 100%, 0.23 min (3.5minute LC-MS method), m/z=170.9, ¹H NMR (500 MHz, Chloroform-d) δ ppm3.79 (s, 2H) 2.31 (d, J=2.84 Hz, 3H) 2.25 (d, J=2.05 Hz, 3H).

Step 3: Synthesis of5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide

A stirred solution of3-[ethyl(oxan-4-yl)amino]-2-methyl-5-[4-(morpholin-4-ylmethyl)phenyl]benzoicacid (100 mg, 0.22 mmol) in anhydrous DMF (4.0 ml) at 0° C. under aballoon of nitrogen, was treated with HATU (99 mg, 0.26 mmol) and DIPEA(75 l, 0.43 mmol) dropwise. The resulting solution was stirred for 10minutes and then treated with3-(aminomethyl)-5-fluoro-4,6-dimethyl-1,2-dihydropyridin-2-one (50%, 81mg, 0.24 mmol). The resulting suspension was stirred at 0° C. for 30minutes and then stirred at room temperature for 18 hours. The reactionmixture was partitioned between water (20 ml) and CH₂Cl₂ (15 ml). Thelayers were separated and the aqueous phase was extracted with CH₂Cl₂(3×15 ml). The combined organics were washed with a saturated solutionof NaHCO₃ (aq) (40 ml), water (2×25 ml), brine (20 ml), dried (MgSO₄),filtered and concentrated in-vacuo. The crude residue was purified byflash column chromatography (10 g SNAP cartridge, Isolera, 0-6%MeOH/CH₂Cl₂) and then dissolved in a mixture of EtOAc (40 ml) and CH₂Cl₂(10 ml), and washed with water (6×30 ml), brine (2×30 ml), dried(MgSO₄), filtered and concentrated in-vacuo. The solid was thoroughlydried in-vacuo at 40° C. for 40 hours to give5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide(93 mg, 73%) as a powdery white solid. LC-MS 100%, 2.76 min (7 minuteLC-MS method), m/z=591.2; ¹H NMR (500 MHz, Chloroform-d) δ 11.79 (s,1H), 7.44 (d, J=8.1 Hz, 2H), 7.34 (d, J=8.4 Hz, 3H), 7.08 (t, J=6.0 Hz,1H), 4.56 (d, J=6.0 Hz, 2H), 3.95 (d, J=11.2 Hz, 2H), 3.76-3.66 (m, 4H),3.51 (s, 2H), 3.31 (td, J=11.3, 2.7 Hz, 2H), 3.10 (q, J=7.0 Hz, 2H),3.00 (tt, J=9.6, 4.6 Hz, 1H), 2.45 (s, 4H), 2.43 (d, J=1.8 Hz, 3H), 2.34(s, 3H), 2.13 (d, J=2.7 Hz, 3H), 1.74-1.62 (m, 4H), 0.89 (t, J=7.0 Hz,3H). One proton assumed to be coincident with a solvent peak.

Example 180:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(1-methyl-1H-imidazol-4-yl)benzamide

Compound 180 was prepared with the method similar to that described inExample 169.

Analytical Data: LCMS: 519.55 (M+1)⁺; HPLC: 89.93% (@ 210-370 nm)(R_(t); 3.676; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.50 (bs, 1H), 9.77 (bs, 1H), 9.11 (s, 1H), 8.20-8.17 (m, 2H), 7.60 (s,1H), 7.38 (s, 1H), 5.87 (s, 1H), 4.30 (d, 2H, J=5.2 Hz), 3.87 (s, 3H),3.11 (m, 3H), 2.68-2.67 (m, 6H), 2.72-2.64 (m, 1H), 2.22 (s, 6H), 2.11(s, 3H), 1.99-1.87 (m, 4H), 1.48-1.40 (m, 4H), 0.81 (t, 3H, J=6.8 Hz).

Example 181:4′-(azetidine-1-carbonyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methyl-[1,1′-biphenyl]-3-carboxamide

Analytical Data: LCMS: 598.60 (M+1)⁺; HPLC: 94.88% (@ 210-370 nm)(R_(t); 3.823; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.44 (bs, 1H), 9.56 (bs, 1H), 8.21 (m, 1H), 8.05-7.96 (m, 4H), 7.54(bs, 1H), 7.38 (bs, 1H), 5.87 (s, 1H), 4.82-4.81 (m, 2H), 4.31 (d, 2H,J=4.8 Hz), 3.69 (t, 2H, J=5.6 Hz), 3.17-3.14 (m, 3H), 2.77 (bs, 1H),2.69-2.68 (m, 6H), 2.26 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.97-1.91(m, 4H), 1.46-1.44 (m, 4H), 0.85 (t, 3H, J=6.8 Hz). 2H merged in solventpeak.

Example 182:N3-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-N4′-(3-hydroxypropyl)-4-methyl-[1,1′-biphenyl]-3,4′-dicarboxamide

Analytical Data: LCMS: 617.70 (M+1)⁺; HPLC: 93.27% (@ 210-370 nm)(R_(t); 4.009; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (bs, 1H), 9.47 (bs, 1H), 8.48 (m, 1H), 8.23 (bs, 1H), 7.93-7.73(m, 4H), 7.47 (bs, 1H), 7.31 (bs, 1H), 5.87 (s, 1H), 4.31-4.30 (m, 2H),3.47 (t, 2H, J=6 Hz), 3.34-3.33 (m, 2H), 3.13 (bs, 3H), 2.69-2.68 (m,6H), 2.26 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.96 (m, 4H), 1.69 (t,2H, J=6.6 Hz), 1.45 (m, 4H), 0.85 (t, 3H). 1H merged in solvent peak.

Example 183:3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)benzamide

Analytical Data: LCMS: 601.65 (M+1)⁺; HPLC: [99.85% (@ 210 nm-370 nm)(R_(t); 4.256; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.48 (bs, 1H), 9.84 (bs, 1H), 8.47 (bs, 1H), 8.17 (bs, 1H), 7.94 (s,1H), 7.41 (m, 2H), 7.04 (d, 1H, J=8 Hz), 5.89 (s, 1H), 4.44 (d, 2H, J=12Hz), 4.30 (s, 2H), 3.84 (bs, 2H), 3.52 (d, 2H, J=9 Hz), 3.12-3.24 (m, 8Hz), 2.85 (s, 3H), 2.25 (s, 3H), 2.11 (s, 3H), 1.54-1.65 (m, 6H),0.84-0.94 (m, 6H). 3 Protons merged in solvent peak.

Example 184: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide

A solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide(0.7 g, 1.38 mmol), respective boronate ester (0.601 g, 2.08 mmol), andtetrakis (0.160 g, 0.138 mmol) in dioxane (10 mL) was purged with argonfor 10 min. To this, aq. Na₂CO₃ (0.529 g, 4.99 mmol, 2 mL) was added andagain degassed for 10 min. Reaction mixture was heated at 100° C. for 16h. On completion, it was concentrated to obtain crude material which wascolumn purified to afford the title compound (0.50 g, 61.5%). AnalyticalData: LCMS: 587.55 (M+1)⁺; HPLC: 97.87% (@ 210-370 nm) (R_(t); 4.217;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase: A; 0.05% TFAin water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30OC; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.45 (bs, 1H),8.38 (d, 1H, J=2.4 Hz), 8.14 (t, 1H, J=4.4 Hz), 7.78 (dd, 1H, J=2.4, 9.2Hz), 7.35 (d, 1H, J=1.2 Hz), 7.15 (d, 1H, J=1.2 Hz), 6.85 (d, 1H, J=8.4Hz), 5.88 (s, 1H), 4.29 (d, 2H, J=4.8 Hz), 3.82 (d, 2H, J=10 Hz), 3.43(t, 4H, J=5.2 Hz), 3.24 (t, 2H, J=11.2 Hz), 3.10-2.98 (m, 3H), 2.78 (t,4H, J=4.8 Hz), 2.22 (s, 3H), 2.11 (s, 3H), 1.67-1.47 (m, 6H), 0.93 (t,3H, J=7.2 Hz), 0.81 (t, 3H, J=6.8 Hz).

Example 185: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide

A solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.5 g, 0.99 mmol),1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)piperazine(0.43 g, 1.48 mmol), and tetrakis (0.114 g, 0.09 mmol) in dioxane (7 mL)was purged with argon for 10 min. To this, aq. Na₂CO₃ (0.377 g, 3.5mmol, 2 mL) was added and again degassed for 10 min. Reaction mixturewas heated at 100° C. for 16 h. On completion, it was concentrated toobtain crude material which was column purified to afford the titlecompound (0.35 g, 60.13%).

Analytical Data: LCMS: 586.36 (M+1)⁺; HPLC: 97.03% (@ 210-370 nm)(R_(t); 4.10; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.45 (bs, 1H), 8.37 (bs, 1H), 8.17 (bs, 1H), 7.78 (d, 1H, J=7.6 Hz),7.35 (s, 1H), 7.15 (s, 1H), 6.85 (d, 1H, J=8.8 Hz), 5.99 (s, 1H), 4.34(d, 2H, J=4 Hz), 3.83-3.81 (m, 2H), 3.42 (bs, 4H), 3.27-3.21 (m, 3H),3.02-3.01 (m, 3H), 2.77 (bs, 4H), 2.22 (s, 3H), 2.13 (s, 3H), 1.67-1.49(m, 4H), 1.13 (s, 3H), 1.12 (s, 3H), 0.81 (t, 3H, J=6.4 Hz).

Example 186: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)benzamide

Step 1: Synthesis of4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile

To a stirred solution of 2-cyanoacetamide (4.1 g, 49 mmol) and t-BuOK(4.9 g, 44.6 mmol) in DMSO at 0° C., 5-methylhex-3-en-2-one (5 g, 44.6mmol) was added and stirred for 30 min. Additional t-BuOK (15 g, 133.9mmol) was added to reaction mixture and stirred at room temperatureunder for further 1 h. On completion, the reaction mixture was dilutedwith water (50 mL) and slowly acidified with 4N HCl. The precipitate wasfiltered and washed with water and dried to get the compound B (2.2 g,28.2%).

Step 2: Synthesis of3-(aminomethyl)-4-isopropyl-6-methylpyridin-2(1H)-one

To a solution of cyano compound B (2.2 g, 12.5 mmol) in methanol and aq.ammonia solution (10 mL, 4:1), catalytic amount of Raney Nickel wasadded. Reaction mass was stirred at room temperature under hydrogenpressure (balloon pressure) for 4 h. On completion of reaction, it wasfiltered through celite bed and filtrate was concentrated under reducepressure to afford the title compound (2 g, 91%).

Step 3: Synthesis of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoic acid(2.0 g, 0.0058 mol) was dissolved in DMSO (20 mL) and3-(aminomethyl)-4-isopropyl-6-methylpyridin-2(1H)-one (2.1 g, 11.7 mmol)and triethyl amine (0.585 g, 5.8 mmol) was added to it. The reactionmixture was stirred at room temperature for 15 min before PyBOP (4.5 g,8.7 mmol) was added to it and stirring was continued for overnight.After completion of the reaction, the reaction mixture was poured intoice, extracted with 10% MeOH/DCM. Combined organic layers were dried,concentrated to obtain crude; which then purified by solvent washings toafford the title compound (2.0 g, 68.9%).

Step 4: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)benzamide

A solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.3 g, 0.99 mmol), respective boronic acid pinacol ester (0.216 g,0.715 mmol), and tetrakis (0.068 g, 0.0596 mmol) in dioxane (10 mL) waspurged with argon for 10 min. To this, aq. Na₂CO₃ (0.227 g, 2.14 mmol, 2mL) was added and again degassed for 10 min. Reaction mixture was heatedat 100° C. for 16 h. On completion, it was concentrated to obtain crudematerial which was purified using prep. HPLC to afford the titlecompound as a TFA salt (0.12 g, 33.6%).

Analytical Data of TFA Salt: MS: 601.55 (M+1)+. HPLC: 96.78% (@ 210-370nm) (R_(t); 4.197; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; MobilePhase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% Bin 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz)δ 11.49 (bs, 1H), 9.94 (bs, 1H), 8.493 (d, 1H, 6 Hz), 7.957 (bs, 1H),7.65-7.258 (m, 3H), 7.056 (d, 1H, 8.4 Hz), 6.014 (s, 1H), 4.46 (d, 2H,12.8 Hz), 4.349 (d, 2H, 4.8 Hz), 3.849 (d, 2H, 7.2 Hz), 3.530 (d, 2H,10.8 Hz), 3.28-3.075 (m, 10H), 2.85 (s, 3H), 2.26 (bs, 3H), 2.14 (s,3H), 1.64 (bs, 2H), 1.56 (bs, 2H),1.14 (s, 3H), 1.12 (s, 3H), 0.845 (t,3H, 7.6 Hz).

Example 187: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)pyridin-3-yl)benzamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide(0.1 g, 0.179 mmol) and 1-methylpiperidin-4-one (0.04 g, 0.358 mmol) indichloroethane (2 mL), acetic acid (0.07 mL, 1.07 mmol) was added andreaction stirred at room temperature for 15 min. Then sodiumtriacetoxyborohydride (0.113 g, 0.53 mmol) was added at 0° C. andreaction stirred overnight at room temperature. On completion, reactionwas quenched with aqueous sodium bicarbonate, organic phase wasseparated and aqueous phase was extracted with dichloromethane. Combinedorganic layers were dried over sodium sulphate and concentrated underreduced pressure to give crude material was purified by prep. HPLC toafford the title compound as a TFA salt (0.08 g, 22.72%).

Analytical Data of TFA salt: ESMS: 656.41 (M+1)⁺; HPLC: [97.76% (@ 210nm-370 nm) (R_(t); 3.667; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30 OC; Flow rate: 1.4 mL/min.; Gradient: 5% B to95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.49 (bs, 1H), 10.33 (bs, 1H), 9.86 (bs, 1H), 8.49 (bs, 1H),8.20 (bs, 1H), 7.96 (bs, 1H), 7.24-7.39 (m, 2H), 7.07 (d, 1H, J=9 Hz),5.87 (s, 1H), 4.47 (bs, 2H), 4.28 (d, 2H, J=4 Hz), 3.84 (s, 2H), 3.60(d, 5H, J=11 Hz), 3.16-3.28 (m, 7H), 2.99 (bs, 2H), 2.79 (s, 3H),2.11-2.25 (m, 11H), 1.87-1.90 (m, 2H), 1.56-1.64 (m, 3H), 0.85 (s, 3H).

Example 188: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)pyridin-3-yl)benzamide

To a stirred solution of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide (0.3 g, 0.51 mmol) and 1-methylpiperidin-4-one (0.086 g, 0.76mmol) in dichloroethane (5 mL), acetic acid (0.18 g, 3.06 mmol) wasadded and reaction stirred at room temperature for 20 minutes. Thensodium triacetoxyborohydride (0.33 g, 1.55 mmol) was added at 0° C. andreaction stirred at room temperature for 2 h. On completion, reactionwas quenched with aqueous sodium bicarbonate, organic phase wasseparated and aqueous phase was extracted with dichloromethane. Combinedorganic layers were dried over sodium sulphate and concentrated underreduced pressure to give crude material was purified by prep. HPLC toafford the title compound (0.12 g, 34.38%).

Analytical Data: LCMS: 683.45 (M+1)⁺; HPLC: 98.65% (@ 210-370 nm)(R_(t); 4.04; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.47 (bs, 1H), 10.24 (bs, 1H), 9.79 (bs, 1H), 8.46 (bs, 1H), 8.19 (bs,1H), 7.92 (bs, 1H), 7.38 (bs, 1H), 7.19 (bs, 1H), 7.06 (d, 1H, J=9.2Hz), 6.0 (s, 1H), 4.47 (bs, 2H), 4.34 (d, 2H, J=7.6 Hz), 3.83 (d, 2HJ=8.8 Hz), 3.6 (d, 4H, J=12 Hz), 3.43 (m, 1H), 3.27-3.16 (m, 8H),2.99-2.97 (m, 3H), 2.79 (s, 3H), 2.37-2.33 (m, 3H), 2.24 (bs, 3H), 2.13(s, 3H), 1.90-1.82 (m, 2H), 1.64-1.53 (m, 4H), 1.13 (s, 3H), 1.12 (s,3H), 0.83 (t, 3H, J=6.8 Hz)

Exmaple 189: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-5-(6-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)pyridin-3-yl)benzamide

To a stirred solution of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide (0.45 g, 0.76 mmol) and 1-methylpiperidin-4-one (0.173 g, 1.53mmol) in dichloroethane (10 mL), acetic acid (0.276 g, 4.6 mmol) wasadded and reaction stirred at room temperature for 20 minutes. Thensodium triacetoxyborohydride (0.488 g, 2.3 mmol) was added at 0° C. andreaction stirred at room temperature for 2 h. On completion, reactionwas quenched with aqueous sodium bicarbonate, organic phase wasseparated and aqueous phase was extracted with dichloromethane. Combinedorganic layers were dried over sodium sulphate and concentrated underreduced pressure to give crude material was purified by columnchromatography to afford the title compound (0.215 g, 41%).

Analytical Data: LCMS: 684.45 (M+1)⁺; HPLC: 93.41% (@ 210 nm-370 nm)(R_(t); 4.140; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (bs, 1H), 8.38 (s, 1H), 8.13 (bs, 1H), 7.78 (d, 1H, J=9 Hz), 7.35(s, 1H), 7.15 (s, 1H), 6.8 (d, 1H, J=9), 5.88 (s, 1H), 4.28 (d, 2H, J=4Hz), 3.82 (d, 2H, 10 Hz), 3.49 (s, 4H), 3.24 (t, 2H, J=11 Hz), 3.0-3.08(m, 3H), 2.78 (d, 2H, J=10 Hz), 2.56 (s, 4H), 2.22 (s, 3H), 2.13 (s,1H), 2.11 (s, 1H), 1.57-1.86 (m, 6H), 1.46-1.55 (m, 6H), 0.91 (t, 3H,J=8 Hz), 0.81 (t, 3H, J=6 Hz).

Example 190: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

A solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.4 g, 0.793 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinaldehyde (0.28 g,1.19 mmol) and tetrakis (0.091 g, 0.079 mmol) in dioxane (5 mL) waspurged with argon for 10 min. To this, aq. Na₂CO₃ (0.301 g, 2.83 mmol)was added and again degassed for 10 min. Reaction mixture was heated at100° C. for 16 h. On completion, it was concentrated to obtain crudematerial which was column purified to afford the title compound (0.28 g,66.50%).

Step 2: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

To a stirred solution of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.28 g, 0.528 mmol) and morpholine (0.07 g, 0.79 mmol) indichloroethane (3 mL), acetic acid (0.19 g, 3.16 mmol) was added andreaction stirred at room temperature for 20 minutes. Then sodiumtriacetoxyborohydride (0.33 g, 1.55 mmol) was added at 0° C. andreaction stirred at room temperature for 2 h. On completion, reactionwas quenched with aqueous sodium bicarbonate, organic phase wasseparated and aqueous phase was extracted with dichloromethane. Combinedorganic layers were dried over sodium sulphate and concentrated underreduced pressure to give crude material was purified by prep. HPLC toafford the title compound (0.12 g, 38.70%).

Analytical Data of TFA salt: LCMS: 601.36 (M+1)⁺; HPLC: 95.48% (@210-370 nm) (R_(t); 4.28; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ;Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.Vol: 10 μL, Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% Bto 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆,400 MHz) δ 11.45 (bs, 1H), 10.45 (bs, 1H), 8.95 (s, 1H), 8.23-8.21 (m,2H), 7.62-7.52 (m, 2H), 7.34 (bs, 1H), 6.01 (s, 1H), 4.55 (s, 2H), 4.35(d, 2H, J=5.2 Hz), 3.84 (bs, 6H), 3.29-3.13 (m, 8H), 2.27 (s, 3H), 2.13(s, 3H), 1.66-1.56 (m, 4H), 1.13 (s, 3H), 1.12 (s, 3H), 0.83 (t, 3H,J=6.8). 2H protons merged in solvent peaks.

Example 191: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide

A solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide(0.5 g, 0.99 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinaldehyde (0.346 g,1.48 mmol), and tetrakis (0.114 g, 0.99 mmol) in dioxane (10 mL) waspurged with argon for 10 min. To this, aq. Na₂CO₃ (0.378 g, 3.56 mmol,1.8 mL) was added and again degassed for 10 min. Reaction mixture washeated at 100° C. for 16 h. On completion, it was concentrated to obtaincrude material which was column purified to afford the title compound(0.40 g, 76.0%).

Step 2: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

To a stirred solution of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-formylpyridin-3-yl)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide(0.315 g, 0.59 mmol) in EDC (8 mL) at 0° C., was added morpholine (0.1g, 1.18 mmol) and stirred at rt for 10 min. NaBH(OAc)₃ (0.377 g, 1.78mmol) was then added and stirred for 16 h. On completion, reaction wasquenched with water. MeOH (8 mL) was added and layers were separated andextracted with 10% MeOH in DCM, and purified on column chromatography toafford the title compound (0.2 g, 56%).

Analytical Data: LCMS: 602.60 (M+1)⁺; HPLC: 98.12% (@ 210 nm-370 nm)(R_(t); 4.374; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.48 (s, 1H), 8.75 (s, 1H), 8.19 (t, 1H, J=4 Hz), 7.99-8.02 (m, 1H),7.49 (t, 2H, J=8 Hz), 7.26 (s, 1H), 5.88 (s, 1H), 4.29 (d, 2H, J=4 Hz),3.82 (d, 2H, J=10 Hz), 3.59-3.61 (m, 6H), 3.24 (t, 2H, J=12 Hz),2.99-3.10 (m, 3H), 2.42 (s, 4H), 2.25 (s, 3H), 2.11 (s, 3H), 1.48-1.67(m, 6H), 0.926 (t, 3H, J=8 Hz), 0.824 (t, 3H, J=7 Hz).

Example 192:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-(4-(dimethylamino)piperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

Step 1: Synthesis of tert-butyl(1-(5-(3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylphenyl)pyridin-2-yl)piperidin-4-yl)carbamate (9)

To a stirred solution of5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(0.35 g, 0.736 mmol) and respective boronic acid pinacol ester (0.35 g,0.88 mmol) in dioxane (5 mL), Na₂CO₃ (0.28 g, 2.65 mmol) was added andsolution was purged with argon for 15 min. Then tetrakis (0.085 g, 0.073mmol) was added and argon was purged again for 10 min. Reaction mass washeated at 100° C. for 4 h. On completion, reaction mixture was dilutedwith water and extracted with 10% MeOH/DCM. Combined organic layers weredried over Na₂SO₄ and solvent removed under reduced pressure andpurified on column chromatography to afford the title compound (0.39 g,79%)

Step 2: Synthesis of5-(6-(4-aminopiperidin-1-yl)pyridin-3-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(10)

To a stirred solution of tert-butyl(1-(5-(3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylphenyl)pyridin-2-yl)piperidin-4-yl)carbamate (0.39 g, 0.058 mmol) in DCM (4 mL) at 0° C.,TFA (2 mL) was added and reaction was stirred for 1 h at roomtemperature. After completion, reaction was concentrated to dryness. Theresidue was then basified with aqueous sat. bicarbonate solution (30 mL)till pH 8 and aqueous layer extracted with 20% methanol in DCM (50mL×4). Combined organic phase was dried over Na₂SO₄ and solvent removedunder reduced pressure to afford the title compound (0.3 g, 90.63%)which was used as such for next reaction.

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(6-(4-(dimethylamino)piperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide

To a stirred solution of5-(6-(4-aminopiperidin-1-yl)pyridin-3-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide(0.3 g, 0.524 mmol) in DCM (3 mL) at 0° C., was added 37-41% aq.formalin solution (0.277 g, 1.31 mmol) and stirred rt for 10 min.NaBH(OAc)₃ (0.277 g, 1.31 mmol) was then added and stirred for 2 h. Oncompletion, reaction was quenched with water. MeOH (10 mL) was added andlayers were separated and extracted with 10% MeOH in DCM, and columnpurified to afford the title compound (0.12 g, 38%).

Analytical Data: LCMS: 602.00 (M+1)⁺; HPLC: 97.22% (@ 210-370 nm)(R_(t); 3.757; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (bs, 1H), 8.38 (s, 1H), 8.15 (t, 1H), 7.78 (d, 1H, J=8.4 Hz), 7.35(s, 1H), 7.16 (s, 1H), 6.90 (d, 1H, J=8.8 Hz), 5.85 (s, 1H), 4.35 (d,2H, J=13.2 Hz), 4.28 (d, 2H, J=4 Hz), 3.82 (d, 2H, J=10 Hz), 3.30-3.20(m, 2H), 3.10-3.00 (m, 3H), 2.90-2.80 (m, 2H), 2.28 (s, 6H), 2.22 (s,3H), 2.20 (s, 3H), 2.10 (s, 3H), 1.90-1.80 (m, 3H), 1.70-1.60 (m, 2H),1.60-1.50 (m, 2H), 1.40-1.30 (m, 2H), 0.82 (t, 3H, J=6.4 Hz).

Example 193: Synthesis of5-(6-(4-(dimethylamino)piperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

Synthesis of tert-butyl(1-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methylphenyl)pyridin-2-yl)piperidin-4-yl)carbamate

A solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.35 g, 0.69 mmol), tert-butyl(1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)piperidin-4-yl)carbamate(0.33 g, 0.83 mmol), and tetrakis (0.079 g, 0.069 mmol) in dioxane (5mL) was purged with argon for 10 min. To this, aq. Na₂CO₃ (0.263 g, 2.48mmol, 2 mL) was added and again degassed for 10 min. Reaction mixturewas heated at 100° C. for 16 h. On completion, it was concentrated toobtain crude material which was column purified to afford the titlecompound (0.31 g, 63%).

Synthesis of5-(6-(4-aminopiperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide

tert-butyl(1-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methylphenyl)pyridin-2-yl)piperidin-4-yl)carbamate(0.31 g, 0.44 mmol) was taken in DCM (5 mL) and TFA (1 mL) was added toit and stirred at rt for 2 h. After completion of reaction, solvent wasremoved under reduced pressure and saturated NaHCO₃ solution was addedto it. Extraction was carried out using 10% MeOH/DCM; the combinedorganic layers were washed with water and brine; dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure to give thetitle compound (0.26 g, 98.11%)

Synthesis of5-(6-(4-(dimethylamino)piperidin-1-yl)pyridin-3-yl)-3-(ethyl-(tetrahydro-2H-pyran-4-yl)-amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-2-methylbenzamide

To a stirred solution of5-(6-(4-aminopiperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide(0.26 g, 0.43 mmol) in DCM (4 mL), formalin (0.045 g, 1.51 mmol) wasadded and reaction stirred at 0° C. of for 10 minutes. Then sodiumtriacetoxyborohydride (0.23 g, 1.08 mmol) was added at 0° C. andreaction stirred for 1 h. On completion, water was added to the reactionmass and extraction was carried out using DCM. Combined organic layerswere washed with bicarbonate solution, dried over sodium sulphate andconcentrated under reduced pressure to give crude material which thenpurified by solvent washings to give the title compound (0.17 g, 62%).

Analytical Data: LCMS: 629.70 (M+1)⁺; HPLC: 97.74% (@ 210-370 nm)(R_(t); 4.176; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (s, 1H), 10.33 (bs, 1H), 8.39 (d, 1H), 8.16 (t, 1H), 7.81 (d, 1H,J=6.8 Hz), 7.35 (s, 1H), 7.16 (s, 1H), 6.97 (d, 1H, J=9.2 Hz), 5.99 (s,1H), 4.50 (d, 2H, J=12.8 Hz), 4.34 (d, 2H, J=4.4 Hz), 3.82 (d, 2H, J=9.6Hz), 3.39 (m, 1H), 3.24 (m, 3H), 3.10-3.00 (m, 3H), 2.90-2.80 (m, 2H),2.69 (s, 6H), 2.22 (s, 3H), 2.13 (s, 3H), 2.10-2.05 (m, 2H), 1.70-1.60(m, 2H), 1.60-1.45 (m, 4H), 1.13 (d, 6H, J=6.4 Hz), 0.82 (t, 3H, J=6.8Hz).

Example 194: Synthesis of5-(6-(4-(dimethylamino)piperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide

Synthesis of tert-butyl(1-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-5-(((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)carbamoyl)phenyl)pyridine-2-yl)piperidin-4-yl)carbamate

A solution of5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide(0.5 g, 0.99 mmol), respective boronic acid pinacol ester (0.6 g, 1.48mmol), and tetrakis (0.114 g, 0.99 mmol) in dioxane (7 mL) was purgedwith argon for 10 min. To this, aq. Na₂CO₃ (0.377 g, 3.5 mmol, 2 mL) wasadded and again degassed for 10 min. Reaction mixture was heated at 100°C. for 16 h. On completion, it was concentrated to obtain crude materialwhich was column purified to afford the title compound (0.40 g, 57.47%).

Synthesis of5-(6-(4-aminopiperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide

To a stirred solution of tert-butyl(1-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-5-(((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)carbamoyl)phenyl)pyridine-2-yl)piperidin-4-yl)carbamate (0.4 g, 0.00051 mol) in DCM (10mL) at 0° C., TFA (10 mL) was added and reaction was stirred for 2 h atroom temperature. After completion, reaction was concentrated todryness. The residue was then basified with aqueous sat. bicarbonatesolution (80 mL) till pH 8 and aqueous layer extracted with 20% methanolin DCM (60 mL×4). Combined organic phase was dried over Na₂SO₄ andsolvent removed under reduced pressure to afford the title compound(0.315 g, 92.1%) which was used as such for next reaction.

Synthesis of5-(6-(4-(dimethylamino)piperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide

To a stirred solution of5-(6-(4-aminopiperidin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide(0.315 g, 0.52 mmol) in DCM (8 mL) at 0° C., was added 37-41% aq.formalin solution (0.078 g, 2.6 mmol) and stirred rt for 10 min.NaBH(OAc)₃ (0.275 g, 1.3 mmol) was then added and stirred for 2 h. Oncompletion, reaction was quenched with water. MeOH (8 mL) was added andlayers were separated and extracted with 10% MeOH in DCM, andrecrystallized from ether, acetonitrile and pentane to afford the titlecompound (0.27 g, 82%).

Analytical Data: LCMS: 630.00 (M+1)⁺; HPLC: 98.21% (@ 210-370 nm)(R_(t); 4.155; Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ; Mobile Phase:A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL,Col. Temp.: 30° C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8min, Hold for 1.5 min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ11.46 (s, 1H), 8.37 (d, 1H, J=1.6 Hz), 8.13 (t, 1H, J=4.4 Hz), 7.76 (dd,1H, J=2.4&9.2 Hz), 7.35 (s, 1H), 7.15 (s, 1H), 6.89 (d, 1H, J=8.8 Hz),5.88 (s, 1H), 4.25-4.35 (m, 4H), 3.82 (d, 2H, J=10 Hz), 3.24 (m, 2H),3.10-3.00 (m, 3H), 2.90-2.80 (m, 2H), 2.35 (m, 1H), 2.22 (s, 3H), 2.18(s, 6H), 2.11 (s, 3H), 1.80 (m, 2H), 1.70-1.60 (m, 2H), 1.60-1.45 (m,4H), 1.40-1.30 (m, 2H), 0.93 (t, 3H, J=7.2 Hz), 0.81 (t, 3H, J=6.8 Hz).

Example 195: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamide

Step 1: Synthesis of5-Fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile

To a stirred solution of6-methyl-2-oxo-4-(propan-2-yl)-1,2-dihydropyridine-3-carbonitrile (225mg, 1.277 mmol) in MeCN (6 mL) was added Selectfluor (620 mg, 1.75mmol). The reaction mixture was stirred at 50° C. for 3 h. After coolingto 23° C., the reaction mixture was concentrated in vacuo. The residuewas purified by column chromatography (50% to 100% EtOAc-heptane toobtain the titled compound (90 mg, 36%). ¹H NMR (400 MHz, CDCl₃) δ ppm3.39 (m, 1H), 2.44 (d, J=3.1 Hz, 3H), 1.41 (dd, J=7.0, 3.1 Hz, 6H); LCMSE-S (M+H)=195.2.

Step 2: Synthesis of3-(Aminomethyl)-5-fluoro-4-isopropyl-6-methylpyridin-2(1H)-one

5-Fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile(100 mg, 0.515 mmol) in 100 mL flask was dissolved in a mixture of MeOH(6 mL) and 2 mL NH_(3aq) (2 mL, 25%). Reduction was conducted usingH-Cube with Raney-Ni as a catalyst at room temperature for 3-4 h. Oncompletion of reaction (monitored by TLC), reaction was concentratedunder reduced pressure to afford titled compound as a grey solid (90 mg,90%). ¹H NMR (400 MHz, CD₃OD) δ ppm 4.04 (s, 2H), 3.22 (m, 1H), 2.24 (d,J=3.4 Hz, 3H), 1.32 (dd, J=7.0, 1.8 Hz, 6H); LCMS E-S (M+H)=199.2.

Step 3: tert-Butyl4-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(methoxycarbonyl)-4-methylphenyl)pyridin-2-yl)piperazine-1-carboxylate

Prepared following the general procedure of Suzuki coupling reaction

¹H-NMR (400 MHz, CDCl₃) δ ppm 8.42 (dd, J=2.4 Hz, 1H), 7.70-7.74 (m,2H), 7.41 (d, J=2.1 Hz, 1H), 6.72 (d, J=8.8 Hz, 1H), 3.97 (m, 2H), 3.93(s, 3H), 3.58 (s, 8H), 3.34 (m, 2H), 3.11 (q, J=7.0 Hz, 2H), 3.02 (m,1H), 2.53 (s, 3H), 1.64-1.76 (m, 4H), 1.50 (s, 9H), 0.91 (t, J=7.0 Hz,3H). MS (ES) (M+H)=539.5.

Step 4: Synthesis of tert-Butyl4-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methylphenyl)pyridin-2-yl)piperazine-1-carboxylate

Hydrolysis of tert-Butyl4-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(methoxycarbonyl)-4-methylphenyl)pyridin-2-yl)piperazine-1-carboxylatefollowing the similar methods for examples described earlier resulted ina crude corresponding carboxylic acid5-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyridin-3-yl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoicacid. This acid was then coupled with3-(aminomethyl)-5-fluoro-4-isopropyl-6-methylpyridin-2(1H)-one followingsimilar methods described earlier. After purification by reverse phaseHPLC (ACN-H₂O containing 0.1% formic acid), the titled compound wasobtained. ¹H-NMR (400 MHz, CD₃OD) δ ppm 8.33 (d, J=2.6 Hz, 1H), 7.81(dd, J=2.6, 8.8 Hz, 1H), 7.41 (d, J=2.0 Hz, 1H), 7.26 (d, J=2.0 Hz, 1H),6.91 (d, J=8.8 Hz, 1H), 4.54 (s, 2H), 3.92 (m, 2H), 3.46-3.56 (m, 9H),3.34 (m, 2H), 3.07-3.18 (m, 3H), 2.33 (s, 3H), 2.24 (d, J=3.2 Hz, 3H),1.74-1.77 (m, 2H), 1.62-1.69 (m, 2H), 1.49 (s, 9H), 1.37 (dd, J=1.6, 6.8Hz, 6H), 0.90 (t, J=6.8 Hz, 3H); MS (ES) (M+H) 705.7.

Step 5: Synthesis of3-(Ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamideformate

To a solution of tert-butyl4-(5-(3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methylphenyl)pyridin-2-yl)piperazine-1-carboxylate(450 mg, 0.639 mmol) in ethanol (4.3 mL) at room temperature was added 4M HCl in dioxane (2 mL, 8.00 mmol). LC/MS after 2 h showed both productand remaining starting material. Additional 4 M HCl in 1,4-dioxane (1.5ml, 6.00 mmol) was added, and LC/MS after total 4 h showed that reactionwas completed. The reaction mixture was concentrated to dryness,azeotroped with toluene-methanol to give crude hydrochloride salt (454mg, 111%). A 125 mg sample of crude hydrochloride salt was purified byreverse phase HPLC/MS (ACN-H₂O, 0.1% formic acid) to give3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamideformate (65 mg) as a colorless glassy film. ¹H-NMR (400 MHz, CD₃OD) δppm 8.35-8.40 (m, 2H), 7.86 (dd, J=2.4, 8.8 Hz, 1H), 7.42 (d, J=1.6 Hz,1H), 7.27 (d, J=1.6 Hz, 1H), 6.99 (d, J=8.8 Hz, 1H), 4.54 (s, 2H), 3.92(m, 2H), 3.84 (m, 4H), 3.54 (m, 1H), 3.30-3.38 (m, 6H), 3.07-3.18 (m,3H), 2.33 (s, 3H), 2.24 (d, J=2.8 Hz, 3H), 1.73-1.76 (m, 2H), 1.62-1.68(m, 2H), 1.37 (d, J=6.8 Hz, 6H), 0.89 (t, J=6.8 Hz, 3H); MS (ES) (M+H)605.6.

Example 196: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)benzamide

To a solution of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(piperazin-1-yl)pyridin-3-yl)benzamidehydrochloride (160 mg, 0.25 mmol) in methanol (2 mL) at 0° C. was added35% solution of formaldehyde in water (0.196 mL, 2.495 mmol). Afterstirring for 20 min, sodium cyanoborohydride (31.4 mg, 0.499 mmol) wasadded. After 1.5 h at 0° C., the reaction was quenched with water (3mL), cooling bath was removed, mixture was stirred for 10 min. Then DCM(10 mL) and saturated aq NaHCO₃ (1 mL) were added. The organic layer wasseparated and the aqueous layer was extracted with DCM (2×15 mL) and thecombined organic extracts were dried over sodium sulfate, filtered andconcentrated. The product was purified by reverse phase HPLC/MS(ACN-H₂O, 0.5% formic acid) to afford 3-(Ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)benzamideformate (31 mg, 0.047 mmol, 19% yield) as a colorless glassy film.

¹H-NMR (400 MHz, CD₃OD) δ ppm 8.42 (br. s, 1H), 8.38 (d, J=2.4 Hz, 1H),7.85 (dd, J=2.8, 8.8 Hz, 1H), 7.42 (d, J=1.6 Hz, 1H), 7.27 (d, J=1.6 Hz,1H), 6.98 (d, J=8.8 Hz, 1H), 4.54 (s, 2H), 3.91 (m, 2H), 3.82 (br. s,4H), 3.54 (m, 1H), 3.31-3.38 (m, 2H), 3.05-3.21 (m, 7H), 2.81 (s, 3H),2.33 (s, 3H), 2.24 (d, J=2.8 Hz, 3H), 1.73-1.76 (m, 2H), 1.58-1.68 (m,2H), 1.37 (dd, J=1.6, 6.8 Hz, 6H), 0.89 (t, J=6.8 Hz, 3H); MS (ES) (M+H)619.7.

Example 197: Synthesis of3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamide

Step 1: Methyl3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzoate

Compound 197 was prepared following the general procedure of Suzukicoupling reaction. ¹H-NMR (400 MHz, CDCl₃) δ ppm 8.77 (dd, J=0.9, 2.4Hz, 1H), 7.84 (dd, J=2.4, 7.9 Hz, 1H), 7.78 (d, J=1.8 Hz, 1H), 7.49 (d,J=7.9 Hz, 1H), 7.45 (d, J=2.1 Hz, 1H), 3.98 (m, 2H), 3.94 (s, 3H),3.75-3.78 (m, 4H), 3.72 (s, 2H), 3.34 (m, 2H), 3.13 (q, J=7.1 Hz, 2H),3.03 (m, 1H), 2.56 (m, 7H), 1.64-1.76 (m, 4H), 0.92 (t, J=7.1 Hz, 3H).MS (ES) (M+H)=454.5.

Step 2:3-(Ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzamideformate

Hydrolysis of methyl3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(6-(morpholinomethyl)pyridin-3-yl)benzoatefollowing similar methods described earlier resulted in thecorresponding carboxylic acid 3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(6-(morpholinomethyl)pyridin-3-yl)benzoic acid. This acid wasthen coupled with3-(aminomethyl)-5-fluoro-4-isopropyl-6-methylpyridin-2(1H)-one followinga similar method described earlier. After purification by reverse phaseHPLC (ACN-H₂O containing 0.1% formic acid), the title compound wasobtained. ¹H-NMR (400 MHz, CD₃OD) δ ppm 8.77 (d, J=1.8 Hz, 1H), 8.25(br. s, 1H), 8.07 (dd, J=2.3, 8.2 Hz, 1H), 7.61 (d, J=8.2 Hz, 1H), 7.52(d, J=1.8 Hz, 1H), 7.37 (d, J=1.8 Hz, 1H), 4.54, (s, 2H), 3.97 (s, 2H),3.91 (m, 2H), 3.78 (m, 4H), 3.55 (m, 1H), 3.35 (m, 2H), 3.08-3.20 (m,3H), 2.81 (m, 4H), 2.36 (s, 3H), 2.24 (d, J=2.9 Hz, 3H), 1.60-1.77 (m,4H), 1.37 (dd, J=1.5, 7.0 Hz, 6H), 0.90 (t, J=6.9 Hz, 3H). MS (ES) (M+H)620.6.

Example 198: Bioassay Protocol and General Methods Protocol forWild-Type and Mutant PRC2 Enzyme Assays

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocyteine (SAH), bicine, KCl,Tween20, dimethylsulfoxide (DMSO) and bovine skin gelatin (BSG) werepurchased from Sigma-Aldrich at the highest level of purity possible.Dithiothreitol (DTT) was purchased from EMD. ³H-SAM was purchased fromAmerican Radiolabeled Chemicals with a specific activity of 80 Ci/mmol.384-well streptavidin Flashplates were purchased from PerkinElmer.

Substrates.

Peptides representative of human histone H3 residues 21-44 containingeither an unmodified lysine 27 (H3K27me0) or dimethylated lysine 27(H3K27me2) were synthesized with a C-terminal G(K-biotin)linker-affinity tag motif and a C-terminal amide cap by 21^(st) CenturyBiochemicals. The peptides were high-performance liquid chromatography(HPLC) purified to greater than 95% purity and confirmed by liquidchromatography mass spectrometry (LC-MS). The sequences are listedbelow.

H3K27me0: (SEQ ID NO: 1) ATKAARKSAPATGGVKKPHRYRPGGK(biotin)-amideH3K27me2: (SEQ ID NO: 2) ATKAARK(me2)SAPATGGVKKPHRYRPGGK(biotin)-amide

Chicken erythrocyte oligonucleosomes were purified from chicken bloodaccording to established procedures.

Recombinant PRC2 Complexes.

Human PRC2 complexes were purified as 4-component enzyme complexesco-expressed in Spodoptera fugiperda (sf9) cells using a baculovirusexpression system. The subunits expressed were wild-type EZH2(NM_004456) or EZH2 Y641F, N, H, S or C mutants generated from thewild-type EZH2 construct, EED (NM_003797), Suz12 (NM_015355) and RbAp48(NM_005610). The EED subunit contained an N-terminal FLAG tag that wasused to purify the entire 4-component complex from sf9 cell lysates. Thepurity of the complexes met or exceeded 95% as determined by SDS-PAGEand Agilent Bioanalyzer analysis. Concentrations of enzyme stockconcentrations (generally 0.3-1.0 mg/mL) was determined using a Bradfordassay against a bovine serum albumin (BSA) standard.

General Procedure for PRC2 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM bicine(pH=7.6), 0.5 mM DTT, 0.005% BSG and 0.002% Tween20, prepared on the dayof use. Compounds in 100% DMSO (1 μL) were spotted into polypropylene384-well V-bottom plates (Greiner) using a Platemate 2×3 outfitted witha 384-channel pipet head (Thermo). DMSO (1 μL) was added to columns 11,12, 23, 24, rows A-H for the maximum signal control, and SAH, a knownproduct and inhibitor of PRC2 (1 μL) was added to columns 11, 12, 23,24, rows I-P for the minimum signal control. A cocktail (40 μL)containing the wild-type PRC2 enzyme and H3K27me0 peptide or any of theY641 mutant enzymes and H3K27me2 peptide was added by Multidrop Combi(Thermo). The compounds were allowed to incubate with PRC2 for 30 min at25° C., then a cocktail (10 μL) containing a mixture of non-radioactiveand ³H-SAM was added to initiate the reaction (final volume=51 L). Inall cases, the final concentrations were as follows: wild-type or mutantPRC2 enzyme was 4 nM, SAH in the minimum signal control wells was 1 mMand the DMSO concentration was 1%. The final concentrations of the restof the components are indicated in Table 2, below. The assays werestopped by the addition of non-radioactive SAM (10 μL) to a finalconcentration of 600 μM, which dilutes the ³H-SAM to a level where itsincorporation into the peptide substrate is no longer detectable. 50 μLof the reaction in the 384-well polypropylene plate was then transferredto a 384-well Flashplate and the biotinylated peptides were allowed tobind to the streptavidin surface for at least 1 h before being washedthree times with 0.1% Tween20 in a Biotek ELx405 plate washer. Theplates were then read in a PerkinElmer TopCount platereader to measurethe quantity of ³H-labeled peptide bound to the Flashplate surface,measured as disintegrations per minute (dpm) or alternatively, referredto as counts per minute (cpm).

TABLE 2 Final concentrations of components for each assay variationbased upon EZH2 identity (wild-type or Y641 mutant EZH2) PRC2 Enzyme(denoted by EZH2 Non-radioactive identity) Peptide (nM) SAM (nM) ³H-SAM(nM) Wild-type 185 1800 150 Y641F 200 850 150 Y641N 200 850 150 Y641H200 1750 250 Y641S 200 1300 200 Y641C 200 3750 250

General Procedure for Wild-Type PRC2 Enzyme Assay on OligonucleosomeSubstrate.

The assays was performed in a buffer consisting of 20 mM bicine(pH=7.6), 0.5 mM DTT, 0.005% BSG, 100 mM KCl and 0.002% Tween20,prepared on the day of use. Compounds in 100% DMSO (1 μL) were spottedinto polypropylene 384-well V-bottom plates (Greiner) using a Platemate2×3 outfitted with a 384-channel pipet head (Thermo). DMSO (1 μL) wasadded to columns 11, 12, 23, 24, rows A-H for the maximum signalcontrol, and SAH, a known product and inhibitor of PRC2 (1 μL) was addedto columns 11, 12, 23, 24, rows I-P for the minimum signal control. Acocktail (40 μL) containing the wild-type PRC2 enzyme and chickenerythrocyte oligonucleosome was added by Multidrop Combi (Thermo). Thecompounds were allowed to incubate with PRC2 for 30 min at 25° C., thena cocktail (10 μL) containing a mixture of non-radioactive and ³H-SAMwas added to initiate the reaction (final volume=51 μL). The finalconcentrations were as follows: wild-type PRC2 enzyme was 4 nM,non-radioactive SAM was 430 nM, ³H-SAM was 120 nM, chicken erythrocyteolignonucleosome was 120 nM, SAH in the minimum signal control wells was1 mM and the DMSO concentration was 1%. The assay was stopped by theaddition of non-radioactive SAM (10 μL) to a final concentration of 600μM, which dilutes the ³H-SAM to a level where its incorporation into thechicken erythrocyte olignonucleosome substrate is no longer detectable.50 μL of the reaction in the 384-well polypropylene plate was thentransferred to a 384-well Flashplate and the chicken erythrocytenucleosomes were immobilized to the surface of the plate, which was thenwashed three times with 0.1% Tween20 in a Biotek ELx405 plate washer.The plates were then read in a PerkinElmer TopCount platereader tomeasure the quantity of ³H-labeled chicken erythrocyte oligonucleosomebound to the Flashplate surface, measured as disintegrations per minute(dpm) or alternatively, referred to as counts per minute (cpm).

%  Inhibition  Calculation

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-parameter  IC₅₀  fit$Y = {{Bottom} + \frac{\left( {{Top} - {Bottom}} \right)}{1 + \left( \frac{X}{{IC}_{50}} \right)^{{Hill}\mspace{14mu} {Coefficient}}}}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

IC₅₀ values for the PRC2 enzyme assays on peptide substrates (e.g., EZH2wild type and Y641F) are presented in Table 3 below.

WSU-DLCL2 Methylation Assay

WSU-DLCL2 suspension cells were purchased from DSMZ (German Collectionof Microorganisms and Cell Cultures, Braunschweig, Germany).RPMI/Glutamax Medium, Penicillin-Streptomycin, Heat Inactivated FetalBovine Serum, and D-PBS were purchased from Life Technologies, GrandIsland, N.Y., USA. Extraction Buffer and Neutralization Buffer (5×) werepurchased from Active Motif, Carlsbad, Calif., USA. Rabbit anti-HistoneH3 antibody was purchased from Abcam, Cambridge, Mass., USA. Rabbitanti-H3K27me3 and HRP-conjugated anti-rabbit-IgG were purchased fromCell Signaling Technology, Danvers, Mass., USA. TMB “Super Sensitive”substrate was sourced from BioFX Laboratories, Owings Mills, Md., USA.IgG-free Bovine Serum Albumin was purchased from Jackson ImmunoResearch,West Grove, Pa., USA. PBS with Tween (10×PBST) was purchased from KPL,Gaithersburg, Md., USA. Sulfuric Acid was purchased from Ricca Chemical,Arlington, Tex., USA. Immulon ELISA plates were purchased from Thermo,Rochester, N.Y., USA. V-bottom cell culture plates were purchased fromCorning Inc., Corning, N.Y., USA. V-bottom polypropylene plates werepurchased from Greiner Bio-One, Monroe, N.C., USA.

WSU-DLCL2 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum and 100units/mL penicillin-streptomycin) and cultured at 37° C. under 5% CO₂.Under assay conditions, cells were incubated in Assay Medium (RPMI 1640supplemented with 20% v/v heat inactivated fetal bovine serum and 100units/mL penicillin-streptomycin) at 37° C. under 5% CO₂ on a plateshaker.

WSU-DLCL2 cells were seeded in assay medium at a concentration of 50,000cells per mL to a 96-well V-bottom cell culture plate with 200 L perwell. Compound (1 μL) from 96 well source plates was added directly toV-bottom cell plate. Plates were incubated on a titer-plate shaker at37° C., 5% CO2 for 96 hours. After four days of incubation, plates werespun at 241×g for five minutes and medium was aspirated gently from eachwell of cell plate without disturbing cell pellet. Pellet wasresuspended in 200 L DPBS and plates were spun again at 241×g for fiveminutes. The supernatant was aspirated and cold (4° C.) Extractionbuffer (100 μL) was added per well. Plates were incubated at 4° C. onorbital shaker for two hours. Plates were spun at 3427×g x 10 minutes.Supernatant (80 μL per well) was transferred to its respective well in96 well V-bottom polypropylene plate. Neutralization Buffer 5× (20 μLper well) was added to V-bottom polypropylene plate containingsupernatant. V-bottom polypropylene plates containing crude histonepreparation (CHP) were incubated on orbital shaker x five minutes. CrudeHistone Preparations were added (2 μL per well) to each respective wellinto duplicate 96 well ELISA plates containing 100 μL Coating Buffer(1×PBS+BSA 0.05% w/v). Plates were sealed and incubated overnight at 4°C. The following day, plates were washed three times with 300 μL perwell 1×BST. Wells were blocked for two hours with 300 L per well ELISADiluent ((PBS (1×) BSA (2% w/v) and Tween20 (0.05% v/v)). Plates werewashed three times with 1×PBST. For the Histone H3 detection plate, 100μL per well were added of anti-Histone-H3 antibody (Abcam, ab1791)diluted 1:10,000 in ELISA Diluent. For H3K27 trimethylation detectionplate, 100 μL per well were added of anti-H3K27me3 diluted 1:2000 inELISA diluent. Plates were incubated for 90 minutes at room temperature.Plates were washed three times with 300 L 1×PBST per well. For HistoneH3 detection, 100 μL of HRP-conjugated anti-rabbit IgG antibody dilutedto 1:6000 in ELISA diluent was added per well. For H3K27me3 detection,100 L of HRP conjugated anti-rabbit IgG antibody diluted to 1:4000 inELISA diluent was added per well. Plates were incubated at roomtemperature for 90 minutes. Plates were washed four times with 1×PBST300 μL per well. TMB substrate 100 μL was added per well. Histone H3plates were incubated for five minutes at room temperature. H3K27me3plates were incubated for 10 minutes at room temperature. The reactionwas stopped with sulfuric acid 1N (100 μL per well). Absorbance for eachplate was read at 450 nm.

First, the ratio for each well was determined by:

$\left( \frac{H\; 3\mspace{14mu} K\; 27{me}\; 3\mspace{14mu} {OD}\; 450\mspace{14mu} {value}}{{Histons}\mspace{14mu} H\; 3\mspace{14mu} {OD}\; 450\mspace{14mu} {value}} \right)$

Each plate included eight control wells of DMSO only treatment (MinimumInhibition) as well as eight control wells for maximum inhibition(Background wells).

The average of the ratio values for each control type was calculated andused to determine the percent inhibition for each test well in theplate. Test compound was serially diluted three-fold in DMSO for a totalof ten test concentrations, beginning at 25 μM. Percent inhibition wasdetermined and IC₅₀ curves were generated using duplicate wells perconcentration of compound. IC₅₀ values for this assay are presented inTable 3 below.

${{Percent}\mspace{14mu} {Inhibition}} = {100 - \left( {\left( \frac{\left. {{Individual}\mspace{14mu} {Test}\mspace{14mu} {Sample}\mspace{14mu} {Ratio}} \right) - \left( {{Background}\mspace{14mu} {Avg}\mspace{14mu} {Ratio}} \right)}{\left( {{Minimum}\mspace{14mu} {Inhibition}\mspace{14mu} {Ratio}} \right) - \left( {{Background}\mspace{14mu} {Average}\mspace{14mu} {Ratio}} \right)} \right)*100} \right)}$

Cell Proliferation Analysis

WSU-DLCL2 suspension cells were purchased from DSMZ (German Collectionof Microorganisms and Cell Cultures, Braunschweig, Germany).RPMI/Glutamax Medium, Penicillin-Streptomycin, Heat Inactivated FetalBovine Serum were purchased from Life Technologies, Grand Island, N.Y.,USA. V-bottom polypropylene 384-well plates were purchased from GreinerBio-One, Monroe, N.C., USA. Cell culture 384-well white opaque plateswere purchased from Perkin Elmer, Waltham, Mass., USA. Cell-Titer Glo®was purchased from Promega Corporation, Madison, Wis., USA. SpectraMaxM5 plate reader was purchased from Molecular Devices LLC, Sunnyvale,Calif., USA.

WSU-DLCL2 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum andcultured at 37° C. under 5% CO₂. Under assay conditions, cells wereincubated in Assay Medium (RPMI 1640 supplemented with 20% v/v heatinactivated fetal bovine serum and 100 units/mL penicillin-streptomycin)at 37° C. under 5% CO₂.

For the assessment of the effect of compounds on the proliferation ofthe WSU-DLCL2 cell line, exponentially growing cells were plated in384-well white opaque plates at a density of 1250 cell/ml in a finalvolume of 50 μl of assay medium. A compound source plate was prepared byperforming triplicate nine-point 3-fold serial dilutions in DMSO,beginning at 10 mM (final top concentration of compound in the assay was20 μM and the DMSO was 0.2%). A 100 nL aliquot from the compound stockplate was added to its respective well in the cell plate. The 100%inhibition control consisted of cells treated with 200 nM finalconcentration of staurosporine and the 0% inhibition control consistedof DMSO treated cells. After addition of compounds, assay plates wereincubated for 6 days at 37° C., 5% CO₂, relative humidity >90% for 6days. Cell viability was measured by quantization of ATP present in thecell cultures, adding 35 μl of Cell Titer Glo® reagent to the cellplates. Luminescence was read in the SpectraMax M5. The concentrationinhibiting cell viability by 50% was determined using a 4-parametric fitof the normalized dose response curves. IC₅₀ values for this assay arealso presented in Table 3 below.

TABLE 3 EZH2 IC50 Y641F WSU prolif WSU ELISA Compound peptide v2 IC₅₀IC₅₀ IC₅₀ No. (μM) (μM) (μM) (μM) 1 0.01745 0.56475 2 0.0549 3 0.24203 40.28847 5 11.21319 6 0.12452 7 28.43469 8 0.13466 9 0.169 10 0.10131 110.01409 1.46188 12 0.07053 13 0.03835 14 0.05688 15 0.1125 16 0.05995 170.02059 18 0.11596 19 0.05865 20 0.03908 21 0.04017 22 0.09501 230.04153 24 0.03473 0.0101 25 0.05556 26 0.0396 0.0273 27 0.02365 0.007212.88863 28 0.03924 29 0.0919 30 0.11932 31 0.045 32 0.06179 35 0.045740.01625 36 0.0149 0.00845 1.54311 37 0.02701 0.05492 38 0.0821 0.0669939 0.01275 0.01432 0.44838 40 0.03107 0.01129 41 0.03176 0.01044 420.04322 0.02206 43 0.02548 0.03009 0.8697 44 0.01299 0.01107 0.369 0.2945 0.07098 0.06219 46 0.0999 0.07546 47 0.03985 0.02028 48 0.096730.07426 49 0.0675 0.04624 50 0.05468 0.0484 51 0.1252 0.1399 52 0.058050.03053 53 0.05837 0.05602 54 0.01367 0.01527 2.40618 55 0.06006 0.0252156 0.03609 0.01737 57 0.03226 0.02333 59 0.01098 0.01513 0.52906 600.23283 0.21286 61 0.04662 0.0414 62 0.17274 0.26915 63 0.0857 0.0682664 0.01055 0.01235 65 0.01132 0.0089 0.15349 66 0.07159 0.04481 0.1673567 0.00653 0.00586 0.11483 68 0.01343 0.02623 0.19913 69 0.00349 0.00260.10184 70 0.03787 0.02958 0.20278 71 0.00415 0.00219 0.18483 72 0.010520.00841 0.27494 73 0.00884 0.00698 0.17821 74 0.00842 0.00632 0.24789 750.00507 0.00348 0.07676 76 0.00374 0.00572 0.09675 77 0.00989 0.005120.15768 78 0.00324 0.00476 2.64294 79 0.00608 0.00778 0.15765 80 0.003110.00388 0.14286 81 0.01054 0.01073 0.40873 82 0.00352 0.00281 0.11923 830.00544 0.00418 0.18335 84 0.01128 0.00612 0.27874 86 0.00499 0.001120.42897 87 0.00568 0.00429 0.15758 0.3332 88 0.00856 0.00591 0.15727 890.00546 0.46186 90 0.00199 0.00361 0.15639 91 0.00315 0.00052 0.13796 920.01169 0.01936 93 0.00258 0.00087 0.10715 94 0.00246 0.00207 0.08649 950.00277 0.00155 0.49957 96 0.01193 0.00899 1.52182 97 0.0034 0.002960.08061 98 0.00582 0.00708 0.35879 99 0.00237 0.00256 0.37993 1000.02155 0.0297 0.43561 101 0.00446 0.01163 0.79789 102 0.02536 0.014840.58584 103 0.00502 0.0082 0.35135 104 0.00963 0.01291 0.33294 1050.00451 0.01065 0.16055 108 0.02337 2.54651 109 0.01921 0.01627 0.68878110 0.00591 0.01239 0.11551 111 0.00766 0.00718 112 0.01831 0.011711.17698 113 0.01883 0.01083 0.35799 114 0.01503 0.01044 0.50615 1150.00783 0.00446 0.21772 116 1.79155 1.2309 117 3.81396 2.30794 >20.0 uM118 0.53042 0.388 4.87739 119 1.5035 0.65543 >20.0 uM 120 0.033040.01566 0.31157 121 0.03614 0.03716 0.29603 122 0.10684 0.07602 0.70354123 0.01159 0.01009 0.29189 124 0.0129 0.00879 0.29994 125 0.024730.02022 0.44695 126 0.01495 0.01178 0.4696 127 0.01177 0.02567 0.3175128 0.00594 0.00695 0.26136 129 0.01782 0.02561 0.29282 130 0.015810.03293 0.63755 131 0.01136 0.02444 0.38733 132 0.00466 0.01225 0.71249133 0.01687 0.02975 0.49827 134 0.01118 0.0189 0.49018 135 0.027570.0484 11.06003 136 0.04262 0.08657 12.29135 137 0.03317 0.02548 1.56152138 0.01173 1.40104 139 0.00707 0.00503 0.30711 140 0.00369 0.004540.37804 141 0.00151 0.00195 0.07815 0.05978 142 1.20523 0.88814 13.37514143 0.00319 0.01274 0.174 144 0.00806 0.00791 0.9863 145 0.00139 0.005530.44891 146 0.01633 0.01575 1.45675 147 0.00344 0.00794 0.19934 1480.01171 0.02295 0.18403 149 0.04316 0.07359 0.63041 150 0.01596 0.05591.46316 151 0.03901 0.03888 152 0.01101 0.02114 0.4062 153 0.004370.00603 0.29683 154 0.02378 0.02848 155 0.01732 0.01753 1.23055 1560.00357 0.00814 0.1114 158 0.0043 0.00509 0.27572 159 0.01524 0.012141.74831 160 0.01211 0.01466 161 0.00438 0.00471 162 0.00574 0.00679 1630.00981 0.00995 4.04577 164 0.01324 0.00514 0.5309 165 0.01133 0.004980.54719 166 0.04563 0.01346 0.80396 167 0.02564 0.00796 0.24542 1680.00995 0.59705 169 0.01238 0.00274 3.26552 170 0.00579 0.00163 0.41075171 0.00397 0.00076 0.3574 0.35597 172 0.00926 0.00421 173 0.012930.00928 0.62489 174 0.0067 0.00289 0.38381 175 0.01195 0.00955 0.28812176 0.01339 0.01035 0.37475 177 >10.0 uM 3.28759 8.1459 178 0.014470.00507 179 0.24404 0.18351 >25.0 uM 180 0.00994 0.00807 181 0.005120.00223 182 0.00666 0.00569 183 0.00466 0.00387 1.52598 184 0.000920.57596 185 0.00338 0.00374 0.41458 186 0.00984 0.52611 188 0.011210.52668 189 0.00164 0.00182 0.1809 190 0.01559 0.53272 191 0.003840.00282 0.37332 192 0.00322 0.34642 193 0.00675 0.0082 0.34313 1940.00462 0.00536 0.64562

Example 199: Derivation of the Lowest Cytotoxic Concentration (LCC)

It is well established that cellular proliferation proceeds through celldivision that results in a doubling of the number of cells afterdivision, relative to the number of cells prior to division. Under afixed set of environmental conditions (e.g., pH, ionic strength,temperature, cell density, medium content of proteins and growthfactors, and the like) cells will proliferate by consecutive doubling(i.e., division) according to the following equation, provided thatsufficient nutrients and other required factors are available.

$\begin{matrix}{N_{t} = {N_{0}2^{\frac{t}{t_{D}}}}} & \left( {A{.1}} \right)\end{matrix}$

where N_(t) is the cell number at a time point (t) after initiation ofthe observation period, No is the cell number at the initiation of theobservation period, t is the time after initiation of the observationperiod and t_(D) is the time interval required for cell doubling, alsoreferred to as the doubling time. Equation A. 1 can be converted intothe more convenient form of an exponential equation in base e, takingadvantage of the equality, 0.693=ln(2).

$\begin{matrix}{N_{t} = {N_{0}^{\frac{0.693t}{t_{D}}}}} & \left( {A{.2}} \right)\end{matrix}$

The rate constant for cell proliferation (k_(p)) is inversely related tothe doubling time as follows.

$\begin{matrix}{k_{p} = \frac{0.693}{t_{D}}} & \left( {A{.3}} \right)\end{matrix}$

Combining equation A.2 and A.3 yields,

N _(t) =N ₀ e ^(k) ^(p) ^(t)  (A.4)

Thus, according to equation A.4 cell number is expected to increaseexponentially with time (FIG. 1 A) during the early period of cellgrowth referred to as log-phase growth. Exponential equations likeequation A.4 can be linearized by taking the natural logarithm of eachside.

ln(N _(t))ln(N ₀)+k _(p) t  (A.5)

Thus a plot of ln(N_(t)) as a function of time is expected to yield anascending straight line with slope equal to k_(p) and y-intercept equalto ln(N₀), as illustrated in FIG. 1 B.

Changes in environmental conditions can result in a change in the rateof cellular proliferation that is quantifiable as changes in theproliferation rate constant k_(p). Among conditions that may result in achange in proliferation rate is the introduction to the system of anantiproliferative compound at the initiation of the observation period(i.e., at t=0). When an antiproliferative compound has an immediateimpact on cell proliferation, one expects that plots of ln(N_(t)) as afunction of time will continue to be linear at all compoundconcentrations, with diminishing values of k_(p) at increasingconcentrations of compound.

Depending on the mechanistic basis of antiproliferative action, somecompounds may not immediately effect a change in proliferation rate.Instead, there may be a period of latency before the impact of thecompound is realized. In such cases a plot of ln(N_(t)) as a function oftime will appear biphasic, and a time point at which the impact of thecompound begins can be identified as the breakpoint between phases (FIG.2). Regardless of whether a compound's impact on proliferation isimmediate or begins after a latency period, the rate constant forproliferation at each compound concentration is best defined by theslope of the ln(N_(t)) vs. time curve from the time point at whichcompound impact begins to the end of the observation period of theexperiment.

A compound applied to growing cells may affect the observedproliferation in one of two general ways: by inhibiting further celldivision (cytostasis) or by cell killing (cytotoxicity). If a compoundis cytostatic, increasing concentration of compound will reduce thevalue of k_(p) until there is no further cell division. At this point,the rate of cell growth, and therefore the value of k_(p), will be zero.If, on the other hand, the compound is cytotoxic, then the value ofk_(p) will be composed of two rate constants: a rate constant forcontinued cell growth in the presence of the compound (k_(g)) and a rateconstant for cell killing by the compound (k_(d)). The overall rateconstant for proliferation at a fixed concentration of compound willthus be the difference between the absolute values of these opposingrate constants.

k _(p) =|k _(g) |−|k _(d)|  (A.6)

At compound concentrations for which the rate of cell growth exceedsthat of cell killing, the value of k_(p) will have a positive value(i.e., k_(p)>0). At compound concentrations for which the rate of cellgrowth is less than that for cell killing, the value of k_(p) will havea negative value (i.e., k_(p)<0) and the cell number will decrease withtime, indicative of robust cytotoxicity. When k_(g) exactly matchesk_(d) then the overall proliferation rate constant, k_(p), will have avalue of zero. We can thus define the lowest cytotoxic concentration(LCC) as that concentration of compound that results in a value of k_(p)equal to zero, because any concentration greater than this will resultin clearly observable cytotoxicity. Nota bene: at concentrations belowthe LCC there is likely to be cell killing occurring, but at a rate thatis less than that of residual cell proliferation. The treatment here isnot intended to define the biological details of compound action.Rather, the goal here is to merely define a practical parameter withwhich to obj ectively quantify the concentration of compound at whichthe rate of cell killing exceeds new cell growth. Indeed, the LCCrepresents a breakpoint or critical concentration above which frankcytotoxicity is observed, rather than a cytotoxic concentration per se.In this regard, the LCC can be viewed similar to other physicalbreakpoint metrics, such as the critical micelle concentration (CMC)used to define the concentration of lipid, detergent or other surfactantspecies above which all molecules incorporate into micellar structures.

Traditionally, the impact of antiproliferative compounds on cell growthhas been most commonly quantified by the IC₅₀ value, which is defined asthat concentration of compound that reduces the rate of cellproliferation to one half that observed in the absence of compound(i.e., for the vehicle or solvent control sample; FIG. 2). The IC₅₀,however, does not allow the investigator to differentiate betweencytostatic and cytotoxic compounds. The LCC, in contrast, readily allowsone to make such a differentiation and to further quantify theconcentration at which the transition to robust cytotoxic behavioroccurs.

If one limits the observation time window to between the start of impact(as defined above and in FIG. 2) and the end of the experiment, then thedata will generally fit well to a linear equation when plotted asln(N_(t)) as a function of time (vide supra). From fits of this type,the value of k_(p) can be determined at each concentration of compoundtested. A replot of the value of k_(p) as a function of compoundconcentration ([I]) will have the form of a descending isotherm, with amaximum value at [I]=0 of k_(max) (defined by the vehicle or solventcontrol sample) and a minimum value at infinite compound concentrationof k_(min) (FIG. 3).

$\begin{matrix}{k_{p} = {\frac{\left( {k_{\max} - k_{\min}} \right)}{1 + \frac{\lbrack I\rbrack}{I_{mid}}} + k_{\min}}} & \left( {A{.7}} \right)\end{matrix}$

where I_(mid) is the concentration of compound yielding a value of k_(p)that is midway between the values of k_(max) and k_(min) (note that thevalue of I_(mid) is not the same as the IC₅₀, except in the case of acomplete and purely cytostatic compound). Thus, fitting the replot datato equation A.7 provides estimates of k_(max), k_(min) and I_(mid). If acompound is cytostatic (as defined here), the value of k_(min) cannot beless than zero. For cytotoxic compounds, k_(min) will be less than zeroand the absolute value of k_(min) will relate directly to theeffectiveness of the compound in killing cells.

The fitted values derived from equation A.7 can also be used todetermine the value of the LCC. By definition, when [I]=LCC, k_(p)=0.Thus, under these conditions equation A.7 becomes.

$\begin{matrix}{0 = {\frac{\left( {k_{\max} - k_{\min}} \right)}{1 + \frac{LCC}{I_{mid}}} + k_{\min}}} & \left( {A{.8}} \right)\end{matrix}$

Algebraic rearrangement of equation A.8 yields an equation for the LCC.

$\begin{matrix}{{LCC} = {I_{mid}\left\lbrack {\left( \frac{k_{\max} - k_{\min}}{- k_{\min}} \right) - 1} \right\rbrack}} & \left( {A{.9}} \right)\end{matrix}$

This analysis is simple to implement with nonlinear curve fittingsoftware and may be applied during cellular assays of compound activitythroughout the drug discovery and development process. In this manner,the LCC may provide a valuable metric for the assessment of compound SAR(structure-activity relationship).

Table 4 below provides LCC and IC₅₀ data for certain compounds of theinvention on WSU-DLCL2 cells.

TABLE 4 11-day LCC Compound (uM) IC50 (uM) No. WSU-DLCL2 WSU-DLCL2 10.68 0.087 2 1.79 0.082 13 0.707 0.018 17 3.32 0.0072 36 0.368 0.011 440.182 0.0093 59 3.15 0.026 65 0.122 0.0018 69 0.0811 0.0062 75 0.05590.00097 87 0.0597 0.0057 67 0.084 0.0028 76 0.165 0.0062 141 0.01530.0023

Example 200: In Vivo Assays Mice

Female Fox Chase SCID® Mice (CB 17/Icr-Prkdc_(scid)/IcrIcoCrl, CharlesRiver Laboratories) or athymic nude mice (Crl:NU(Ncr)-Foxn1_(nu),Charles River Laboratories) were 8 weeks old and had a body-weight (BW)range of 16.0-21.1 g on D1 of the study. The animals were fed ad libitumwater (reverse osmosis 1 ppm Cl) and NIH 31 Modified and Irradiated LabDiet® consisting of 18.0% crude protein, 5.0% crude fat, and 5.0% crudefiber. The mice were housed on irradiated Enrich-o'cobs™ bedding instatic microisolators on a 12-hour light cycle at 20-22° C. (68-72° F.)and 40-60% humidity. All procedures comply with the recommendations ofthe Guide for Care and Use of Laboratory Animals with respect torestraint, husbandry, surgical procedures, feed and fluid regulation,and veterinary care.

Tumor Cell Culture

Human lymphoma cell lines line were obtained from different sources(ATCC, DSMZ), e.g., WSU-DLCL2 obtained from DSMZ. The cell lines weremaintained at Piedmont as suspension cultures in RPMI-1640 mediumcontaining 100 units/mL penicillin G sodium salt, 100 g/mL streptomycin,and 25 g/mL gentamicin. The medium was supplemented with 10% fetalbovine serum and 2 mM glutamine. The cells were cultured in tissueculture flasks in a humidified incubator at 37° C., in an atmosphere of5% CO₂ and 95% air.

In Vivo Tumor Implantation

Human lymphoma cell lines, e.g., WSU-DLCL2 cells, were harvested duringmid-log phase growth, and re-suspended in PBS with 50% Matrigel™ (BDBiosciences). Each mouse received 1×10⁷ cells (0.2 mL cell suspension)subcutaneously in the right flank. Tumors were calipered in twodimensions to monitor growth as the mean volume approached the desired80-120 mm³ range. Tumor size, in mm³, was calculated from:

${{Tumor}\mspace{14mu} {volume}} = \frac{w^{2}l}{2}$

where w=width and l=length, in mm, of the tumor. Tumor weight can beestimated with the assumption that 1 mg is equivalent to 1 mm₃ of tumorvolume. After 10-30 days mice with 108-126 mm³ tumors were sorted intotreatment groups with mean tumor volumes of 117-119 mm³.

Test Articles

Test compounds were stored at room temperature and protected from light.On each treatment day, fresh compound formulations (e.g., formulation ofCompound 44 tri-HCl salt or Compound 87 tri-HCl salt), were prepared bysuspending the powders in 0.5% sodium carboxymethylcellulose (NaCMC) and0.1% Tweeno 80 in deionized water. Compound 141 (free base) wasdissolved in sterile saline and the pH was adjusted to 4.5 with HClfresh every day. The vehicles, 0.5% NaCMC and 0.1% Tween® 80 indeionized water or sterile saline pH 4.5, were used to treat the controlgroups at the same schedules. Formulations were stored away from lightat 4° C. prior to administration. Unless otherwise specified, compoundsreferred to and tested in this experiment were in their specific saltforms mentioned in this paragraph.

Treatment Plan

Mice were treated at compound doses ranging from 12.5-600 mg/kg and atTID (three time a day every 8 h), BID (2 times a day every 12 h) or QD(once a day) schedules for various amounts of days by oral gavage(Compound 44 or 87) or injections via the intraperitoneal route(Compound 141). Each dose was delivered in a volume of 0.2 mL/20 g mouse(10 mL/kg), and adjusted for the last recorded weight of individualanimals. The maximal treatment length was 28 days.

Median Tumor Volume (MTV) and Tumor Growth Inhibition (TGI) Analysis

Treatment efficacy was determined on the last treatment day. MTV(n), themedian tumor volume for the number of animals, n. evaluable on the lastday, was determined for each group. Percent tumor growth inhibition (%TGI) can be defined several ways. First, the difference between theMTV(n) of the designated control group and the MTV(n) of thedrug-treated group is expressed as a percentage of the MTV(n) of thecontrol group:

${\% \mspace{20mu} {TGI}} = {\left( \frac{{{MTV}(n)}_{control} - {{MTV}(n)}_{treated}}{{{MTV}(n)}_{control}} \right)100}$

Another way of calculating % TGI is taking the change of the tumor sizefrom day 1 to day n into account with n being the last treatment day.

${\% \mspace{20mu} {TGI}} = {\left( \frac{{\Delta \; {MTV}_{control}} - {\Delta \; {MTV}_{treated}}}{\Delta \; {MTV}_{control}} \right)100}$Δ MTV_(control) = MTV(n)_(control) − MTV(1)_(control)Δ MTV_(treated) = MTV(n)_(treated) − MTV(1)_(treated)

Toxicity

Animals were weighed daily on Days 1-5, and then twice weekly until thecompletion of the study. The mice were examined frequently for overtsigns of any adverse, treatment related side effects, which weredocumented. Acceptable toxicity for the maximum tolerated dose (MTD) wasdefined as a group mean BW loss of less than 20% during the test, andnot more than 10% mortality due to TR deaths. A death was to beclassified as TR if it was attributable to treatment side effects asevidenced by clinical signs and/or necropsy, or due to unknown causesduring the dosing period. A death was to be classified as NTR if therewas evidence that the death was unrelated to treatment side effects. NTRdeaths during the dosing interval would typically be categorized as NTRa(due to an accident or human error) or NTRm (due to necropsy-confirmedtumor dissemination by invasion and/or metastasis). Orally treatedanimals that die from unknown causes during the dosing period may beclassified as NTRu when group performance does not support a TRclassification and necropsy, to rule out a dosing error, is notfeasible.

Sampling

On days 7 or 28 during the studies mice were sampled in a pre-specifiedfashion to assess target inhibition in tumors. Tumors were harvestedfrom specified mice under RNAse free conditions and bisected. Frozentumor tissue from each animal was snap frozen in liquid N2 andpulverized with a mortar and pestle.

Statistical and Graphical Analyses

All statistical and graphical analyses were performed with Prism 3.03(GraphPad) for Windows. To test statistical significance between thecontrol and treated groups over the whole treatment time course arepeated measures ANOVA test followed by Dunnets multiple comparisonpost test or a 2 way ANOVA test were employed. Prism reports results asnon-significant (ns) at P>0.05, significant (symbolized by “*”) at0.01<P<0.05, very significant (“**”) at 0.001<P<0.01 and extremelysignificant (“***”) at P<0.001.

Histone Extraction

For isolation of histones, 60-90 mg tumor tissue was homogenized in 1.5ml nuclear extraction buffer (10 mM Tris-HCl, 10 mM MgCl2, 25 mM KCl, 1%Triton X-100, 8.6% Sucrose, plus a Roche protease inhibitor tablet1836145) and incubated on ice for 5 minutes. Nuclei were collected bycentrifugation at 600 g for 5 minutes at 4° C. and washed once in PBS.Supernatant was removed and histones extracted for one hour, withvortexing every 15 minutes, with 0.4 N cold sulfuric acid. Extracts wereclarified by centrifugation at 10000 g for 10 minutes at 4° C. andtransferred to a fresh microcentrifuge tube containing 10× volume of icecold acetone. Histones were precipitated at −20° C. for 2hours-overnight, pelleted by centrifugation at 10000 g for 10 minutes,and resuspended in water.

ELISA

Histones were prepared in equivalent concentrations in coating buffer(PBS+0.05% BSA) yielding 0.5 ng/ul of sample, and 100 ul of sample orstandard was added in duplicate to 2 96-well ELISA plates (ThermoLabsystems, Immulon 4HBX #3885). The plates were sealed and incubatedovernight at 4° C. The following day, plates were washed 3× with 300ul/well PBST (PBS+0.05% Tween 20; 10×PBST, KPL #51-14-02) on a Bio Tekplate washer. Plates were blocked with 300 ul/well of diluent (PBS+2%BSA+0.05% Tween 20), incubated at RT for 2 hours, and washed 3× withPBST. All antibodies were diluted in diluent. 100 ul/well ofanti-H3K27me3 (CST #9733, 50% glycerol stock 1:1,000) or anti-total H3(Abcam ab1791, 50% glycerol 1:10,000) was added to each plate. Plateswere incubated for 90 min at RT and washed 3× with PBST. 100 ul/well ofanti-Rb-IgG-HRP (Cell Signaling Technology, 7074) was added 1:2,000 tothe H3K27Me3 plate and 1:6,000 to the H3 plate and incubated for 90 minat RT. Plates were washed 4× with PBST. For detection, 100 ul/well ofTMB substrate (BioFx Laboratories, #TMBS) was added and plates incubatedin the dark at RT for 5 min. Reaction was stopped with 100 ul/well 1NH₂SO₄. Absorbance at 450 nm was read on SpectaMax M5 Microplate reader.

Results:

7 Day PD Study with Compound 87

In order to test whether Compound 87 can modulate the H3K27me3 histonemark in tumors in vivo, WSU-DLCL2 xenograft tumor bearing mice weretreated with Compound 87 at either 200 mg/kg BID or 400 mg/kg QD orvehicle (BID schedule) for 7 days. There were 4 animals per group.Animals were euthanized 3 h after the last dose and tumor was preservedin a frozen state as described above. Following histone extraction thesamples were applied to ELISA assays using antibodies directed againstthe trimethylated state of histone H3K27 (H3K27me3) or total histone H3.Based on these data the ratio of globally methylated to total H3K27 wascalculated. FIG. 4 shows the mean global methylation ratios for allgroups as measures by ELISA and indicates target inhibition ranging fromapp. 62.5% (400 mg/kg QD×7) and 37.5% (200 mg/kg BID×7) compared tovehicle.

28 Day Efficacy Study with Compound 141 in WSU-DLCL2 Xenograft Model

In order to test whether Compound 141 could induce a tumor growthinhibition in vivo WSU-DLCL2 xenograft tumor bearing mice were treatedwith Compound 141 at 12.5, 25 or 50 mg/kg QD for 28 days viaintraperitoneal injection. Tumor volume and body weights were determinedtwice a week. Compound 141 was well tolerated at all doses with minimalbody weight loss. A parallel cohort of mice (n=4 per group) was treatedat the same doses for 7 days, and mice were euthanized on day 7, 3 hafter the last dose for tumor sampling and assessment of targetinhibition. FIG. 5 shows the result of the ELISA measuring globalmethylation of H3K27me3 normalized to total H3. Dose dependent targetinhibition ranging from 39% to 67% compared to vehicle can be observed.

FIG. 6 shows the tumor growth over the treatment course of 28 days forthe groups treated with vehicle or Compound 141.

An effect of administration of the vehicle given via the intraperitonealroute could be observed as tumor growth was slower in the vehicle groupvs. the untreated group. Only the highest dose group of Compound 141 (50mg/kg QD×28) showed tumor growth inhibition compared to the vehiclegroup (33% calculated from day 1, 43% calculated from day 7). The tumorgrowth was not statistically significant compared to vehicle when usinga repeated measures ANOVA followed by Dunnets post test, but theterminal tumor size was significantly smaller in the 50 mg/kg DQ groupcompared to vehicle (2 way ANOVA, Bonferroni post test, p<0.0001).

Efficacy Study with Increasing Doses of Compound 44 in WSU-DLCL2Xenograft Model

In order to test whether compound 44 could induce an anti-tumor effectin vivo WSU-DLCL2 xenograft tumor bearing mice were treated withcompound 44 at 37.5, 75 or 150 mg/kg TID for 28 days. There were 12 miceper group for the efficacy arm of the experiment. A parallel cohort wasdosed for 7 days at the same doses and schedules for assessment oftarget inhibition after 7 days (n=6 per group). FIG. 7 shows the tumorgrowth over the treatment course of 28 days for vehicle and compound 44treated groups. A clear dose dependent tumor growth inhibition could beobserved. Only the highest dose group was statistically significant fromvehicle by repeated measures ANOVA and Dunnett's post test. The tumorgrowth inhibition for the highest dose group was 58% (from day 1) or 73%(from day 7) compared to vehicle.

Histones were extracted from tumors collected after 7 days of dosing(parallel PD cohort) and at the end of the study on day 28 for theefficacy cohort (3 h after the last dose for both cohorts). FIG. 8 showsthat the H3K27me3 methyl mark is modulated with treatment in a dosedependent matter and that there is statistically significant bettertarget inhibition on day 28 compared to day 7 for the highest dose group(150 mg/kg TID).

Efficacy Study with Compound 44 at Different Dose Schedules

To assess whether Compound 44 would lead to tumor growth inhibition atother dosing schedules but TID a WSU-DLCL2 xenograft efficacy study wasperformed where TID, BID and QD schedules were compared side by side.There were 12 animals per group, and mice were treated for 28 days. FIG.9 shows the tumor growth over the treatment course of 28 days forvehicle and Compound 44 treated groups. Tumor growth inhibition could beachieved at all doses and schedules (Table 5 below: summary of tumorgrowth inhibitions induced by different dosing schedules of Compound 44in WSU-DLXC2 xenografts). While only the 150 mg/kg TID and 225 mg/kg BIDwere statistically significant from vehicle by repeated measures ANOVAand Dunnett's post test all terminal tumor sizes in Compound 44 treatedgroups were statistically different from vehicle by 2 way ANOVA andBonferroni post test (p<0.0001).

TABLE 5 Group % TGI from day 1 % TGI from day 7 150 mg/kg TID 73 86 225mg/kg BID 71 80 300 mg/kg BID 57 67 600 mg/kg QD 58 70

On day 28 mice were euthanized and tumors were collected 3 h after thelast dose for assessment of target inhibition. FIG. 10 shows thattreatment with Compound 44 induced similar degrees of target inhibitionfor all doses and schedules.

Example 201: Anti-Cancer Effect of Compound 44 on the KARPAS-422 HumanDiffused Large B-Cell Lymphoma Mouse Xenograft Model

Compound 44 (tri-HCl salt form) was tested for its anti-cancer activityin KARPAS-422 mouse xenograft model, which is a human diffused largeB-Cell lymphoma xenograft model. Unless otherwise specified, Compound 44referred to and tested in this experiment was its tri-HCl salt form. 45female of CAnN.Cg-Foxn1nu/CrlCrlj mice (Charles River LaboratoriesJapan) with KARPAS-422 tumors whose mean tumor volume (TV) reachedapproximately 150 mm³ were selected based on their TVs, and wererandomly divided into five groups. The oral administration of Compound44 (80.5, 161, 322, and 644 mg/kg) or vehicle was started on day 1.Compound 44 was given once daily on day 1 and day 29 and twice dailyeveryday from day 2 to day 28. The administration volume (0.1 mL/10 gbody weight) was calculated from the body weight before administration.The TV and body weight were measured twice a week. The design for thisexperiment is shown in Table 6.

TABLE 6 Dosing Scheme No. of Group Animals Treatment (twice a day) Routeand Schedule 1 9 Vehicle (0.5% Methyl PO; BID × 28 days Cellulose, 0.1%Tween-80) 2 9 80.5 mg/kg Compound 44 PO; BID × 28 days (tri-HCl salt) 39 161 mg/kg Compound 44 PO; BID × 28 days (tri-HCl salt) 4 9 322 mg/kgCompound 44 PO; BID × 28 days (tri-HCl salt) 5 9 644 mg/kg Compound 44PO; bid × 28 days (tri-HCl salt)

TV is calculated from caliper measurements by the formula for the volumeof a prolate ellipsoid (L×W²)/2 where L and W are the respectiveorthogonal length and width measurements (mm).

Data are expressed as the mean+standard deviation (SD). The differencesin TV between the vehicle-treated and Compound 44-treated groups wereanalyzed by a repeated measures analysis of variance (ANOVA) followed bythe Dunnett-type multiple comparison test. A value of P<0.05 (two sided)was considered statistically significant. Statistical analyses wereperformed using the Prism 5 software package version 5.04 (GraphPadSoftware, Inc., CA, USA).

During the treatment period, the dosage of 644 mg/kg resulted in thedeath of two out of nine mice. The maximum tolerated dose was determinedas 322 mg/kg, at which dosage no mortality or no body weight loss wasrecorded, in the study (FIG. 11 and Table 7).

Compound 44 showed significant antitumor effects against a human diffuselarge B cell lymphoma KARPAS-422 xenograft at all of the doses on day 29in a dose dependent manner (FIG. 12). Tumor growth-inhibition effect wasobserved at 80.5 mg/kg. Tumor regressions were observed at 161 and 322mg/kg.

TABLE 7 Effect of Compound 44 on Mouse Mortality Treatment Number twicea day dead/total Vehicle 0/9 Cpd 44 (tri-HCl salt) 0/9 80.5 mg/kg Cpd 44(tri-HCl salt) 0/9 161 mg/kg Cpd 44(tri-HCl salt) 0/9 322 mg/kg Cpd 44(tri-HCl salt) 2/9 644 mg/kg

INCORPORATION BY REFERENCE

The entire disclosure of each of the patent documents and scientificarticles referred to herein is incorporated by reference for allpurposes.

EQUIVALENTS

The invention can be embodied in other specific forms without departingfrom the spirit or essential characteristics thereof. The foregoingembodiments are therefore to be considered in all respects illustrativerather than limiting on the invention described herein. Scope of theinvention is thus indicated by the appended claims rather than by theforegoing description, and all changes that come within the meaning andrange of equivalency of the claims are intended to be embraced therein.

What is claimed is:
 1. A method of treating soft tissue sarcoma,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof:

wherein X₁ is N or CR₁₁; X₂ is N or CR₁₃; Z is NR₇R₈, OR₇, S(O)_(n)R₇,or CR₇R₈R₁₄, in which n is 0, 1, or 2; each of R₁, R₅, R₉, and R₁₀,independently, is H or C₁-C₆ alkyl optionally substituted with one ormore substituents selected from the group consisting of halo, hydroxyl,COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl; each ofR₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is a bond or C₁-C₃alkyl linker optionally substituted with halo, cyano, hydroxyl or C₁-C₆alkoxy, and T₁ is H, halo, hydroxyl, COOH, cyano, or R_(S1), in whichR_(S1) is C₁-C₃ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl,C(O)O—C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, 4 to 12-membered heterocycloalkyl, or5- or 6-membered heteroaryl, and R_(S1) is optionally substituted withone or more substituents selected from the group consisting of halo,hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀aryl, 4 to 12-membered heterocycloalkyl, and 5- or 6-memberedheteroaryl; R₆ is C₆-C₁₀ aryl or 5- or 6-membered heteroaryl, each ofwhich is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is abond or C₁-C₃ alkyl linker optionally substituted with halo, cyano,hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano, —OR_(a),—NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, —C(O)R_(a), —C(O)OR_(a),—C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a), —NR_(b)C(O)OR_(a), —S(O)₂R_(a),—S(O)₂NR_(a)R_(b), or R_(S2), in which each of R_(a), R_(b), and R_(e),independently is H or R_(S3), A⁻ is a pharmaceutically acceptable anion,each of R_(S2) and R_(S3), independently, is C₁-C₆ alkyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, or 5- or6-membered heteroaryl, or R_(a) and R_(b), together with the N atom towhich they are attached, form a 4 to 12-membered heterocycloalkyl ringhaving 0 or 1 additional heteroatom, and each of R_(S2), R_(S3), and the4 to 12-membered heterocycloalkyl ring formed by R_(a) and R_(b), isoptionally substituted with one or more -Q₃-T₃, wherein Q₃ is a bond orC₁-C₃ alkyl linker each optionally substituted with halo, cyano,hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from the group consistingof halo, cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, OR_(d),COOR_(d), —S(O)₂R_(d), —NR_(d)R_(e), and —C(O)NR_(d)R_(e), each of R_(d)and R_(e) independently being H or C₁-C₆ alkyl, or -Q₃-T₃ is oxo; or anytwo neighboring -Q₂-T₂, together with the atoms to which they areattached form a 5- or 6-membered ring optionally containing 1-4heteroatoms selected from N, O and S and optionally substituted with oneor more substituents selected from the group consisting of halo,hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀aryl, 4 to 12-membered heterocycloalkyl, and 5- or 6-memberedheteroaryl; R₇ is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄ alkyl linker, orC₂-C₄ alkenyl linker, each linker optionally substituted with halo,cyano, hydroxyl or C₁-C₆ alkoxy, and T₄ is H, halo, cyano, NR_(f)R_(g),—OR_(f), —C(O)R_(f), —C(O)OR_(f), —C(O)NR_(f)R_(g), —C(O)NR_(f)OR_(g),—NR_(f)C(O)R_(g), —S(O)₂R_(f), or R_(S4), in which each of R_(f) andR_(g), independently is H or R_(S5), each of R_(S4) and R_(S5),independently is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, or 5- or6-membered heteroaryl, and each of R_(S4) and R_(S5) is optionallysubstituted with one or more -Q₅-T₅, wherein Q₅ is a bond, C(O),C(O)NR_(k), NR_(k)C(O), S(O)₂, or C₁-C₃ alkyl linker, R_(k) being H orC₁-C₆ alkyl, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, 5- or6-membered heteroaryl, or S(O)_(q)R_(q) in which q is 0, 1, or 2 andR_(q) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl,C₆-C₁₀ aryl, 4 to 12-membered heterocycloalkyl, or 5- or 6-memberedheteroaryl, and T₅ is optionally substituted with one or moresubstituents selected from the group consisting of halo, C₁-C₆ alkyl,hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-memberedheterocycloalkyl, and 5- or 6-membered heteroaryl except when T₅ is H,halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; each of R₈, R₁₁, R₁₂, andR₁₃, independently, is H, halo, hydroxyl, COOH, cyano, R_(S6), OR_(S6),or COOR_(S6), in which R_(S6) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₃-C₈ cycloalkyl, 4 to 12-membered heterocycloalkyl, amino,mono-C₁-C₆ alkylamino, or di-C₁-C₆ alkylamino, and R_(S6) is optionallysubstituted with one or more substituents selected from the groupconsisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆ alkylamino; or R₇and R₈, together with the N atom to which they are attached, form a 4 to11-membered heterocycloalkyl ring having 0 to 2 additional heteroatoms,or R₇ and R₈, together with the C atom to which they are attached, formC3-C8 cycloalkyl or a 4 to 11-membered heterocycloalkyl ring having 1 to3 heteroatoms, and each of the 4 to 11-membered heterocycloalkyl ringsor C₃-C₈ cycloalkyl formed by R₇ and R₈ is optionally substituted withone or more -Q₆-T₆, wherein Q₆ is a bond, C(O), C(O)NR_(m), NR_(m)C(O),S(O)₂, or C₁-C₃ alkyl linker, R_(m) being H or C₁-C₆ alkyl, and T₆ is H,halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, orS(O)_(p)R_(p) in which p is 0, 1, or 2 and R_(p) is C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-memberedheterocycloalkyl, or 5- or 6-membered heteroaryl, and T₆ is optionallysubstituted with one or more substituents selected from the groupconsisting of halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino,mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀aryl, 4 to 12-membered heterocycloalkyl, and 5- or 6-membered heteroarylexcept when T₆ is H, halo, hydroxyl, or cyano; or -Q₆-T₆ is oxo; and R₁₄is absent, H, or C₁-C₆ alkyl optionally substituted with one or moresubstituents selected from the group consisting of halo, hydroxyl, COOH,C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 12-memberedheterocycloalkyl, and 5- or 6-membered heteroaryl.
 2. The method ofclaim 1, wherein: X₁ is N or CR₁₁; X₂ is N or CR₁₃; Z is NR₇T₈, OR₇,SR₇, or CR₇R₈R₁₄; each of R₁, R₅, R₉, and R₁₀, independently, is H orC₁-C₆ alkyl optionally substituted with one or more substituentsselected from the group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-memberedheterocycloalkyl, and 5- or 6-membered heteroaryl; each of R₂, R₃, andR₄, independently, is -Q₁-T₁, in which Q₁ is a bond or C₁-C₃ alkyllinker optionally substituted with halo, cyano, hydroxyl or C₁-C₆alkoxy, and T₁ is H, halo, hydroxyl, COOH, cyano, or R_(S1), in whichR_(S1) is C₁-C₃ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl,C(O)O—C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, 4 to 7-membered heterocycloalkyl, or 5-or 6-membered heteroaryl, and R_(S1) is optionally substituted with oneor more substituents selected from the group consisting of halo,hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀aryl, 4 to 7-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;R₆ is C₆-C₁₀ aryl or 5- or 6-membered heteroaryl, each of which isoptionally substituted with one or more -Q₂-T₂, wherein Q₂ is a bond orC₁-C₃ alkyl linker optionally substituted with halo, cyano, hydroxyl orC₁-C₆ alkoxy, and T₂ is H, halo, cyano, —OR_(a), —NR_(a)R_(b),—(NR_(a)R_(b)R_(c))⁺A⁻, —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b),—NR_(b)C(O)R_(a), —NR_(b)C(O)OR_(a), —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), orR_(S2), in which each of R_(a), R_(b), and R_(e), independently is H orR_(S3), A⁻ is a pharmaceutically acceptable anion, each of R_(S2) andR_(S3), independently, is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4to 7-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, or R_(a)and R_(b), together with the N atom to which they are attached, form a 4to 7-membered heterocycloalkyl ring having 0 or 1 additional heteroatom,and each of R_(S2), R_(S3), and the 4 to 7-membered heterocycloalkylring formed by R_(a) and R_(b), is optionally substituted with one ormore one or more -Q₃-T₃, wherein Q₃ is a bond or C₁-C₃ alkyl linker eachoptionally substituted with halo, cyano, hydroxyl or C₁-C₆ alkoxy, andT₃ is selected from the group consisting of halo, cyano, C₁-C₆ alkyl,C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, 5- or6-membered heteroaryl, OR_(d), COOR_(d), —S(O)₂R_(d), —NR_(d)R_(e), and—C(O)NR_(d)R_(e), each of R_(d) and R_(e) independently being H or C₁-C₆alkyl, or -Q₃-T₃ is oxo; or any two neighboring -Q₂-T₂, together withthe atoms to which they are attached form a 5- or 6-membered ringoptionally containing 1-4 heteroatoms selected from N, O and S andoptionally substituted with one or more substituents selected from thegroup consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano,C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5- or6-membered heteroaryl; provided that -Q₂-T₂ is not H; R₇ is -Q₄-T₄, inwhich Q₄ is a bond, C₁-C₄ alkyl linker, or C₂-C₄ alkenyl linker, eachlinker optionally substituted with halo, cyano, hydroxyl or C₁-C₆alkoxy, and T₄ is H, halo, cyano, NR_(f)R_(g), —OR_(f), —C(O)R_(f),—C(O)OR_(f), —C(O)NR_(f)R_(g), —C(O)NR_(f)OR_(g), —NR_(f)C(O)R_(g),—S(O)₂R_(f), or R_(S4), in which each of R_(f) and R_(g), independentlyis H or R_(S5), each of R_(S4) and R_(S5), independently is C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to7-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and each ofR_(S4) and R_(S5) is optionally substituted with one or more -Q₅-T₅,wherein Q₅ is a bond, C(O), C(O)NR_(k), NR_(k)C(O), S(O)₂, or C₁-C₃alkyl linker, R_(k) being H or C₁-C₆ alkyl, and T₅ is H, halo, C₁-C₆alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-memberedheterocycloalkyl, 5- or 6-membered heteroaryl, or S(O)_(q)R_(q) in whichq is 0, 1, or 2 and R_(q) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, or 5-or 6-membered heteroaryl, and T₅ is optionally substituted with one ormore substituents selected from the group consisting of halo, C₁-C₆alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-memberedheterocycloalkyl, and 5- or 6-membered heteroaryl except when T₅ is H,halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; provided that R₇ is not H;each of R₈, R₁₁, R₁₂, and R₁₃, independently, is H, halo, hydroxyl,COOH, cyano, R_(S6), OR_(S6), or COOR_(S6), in which R_(S6) is C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, amino, mono-C₁-C₆ alkylamino, ordi-C₁-C₆ alkylamino, and R_(S6) is optionally substituted with one ormore substituents selected from the group consisting of halo, hydroxyl,COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, and di-C₁-C₆ alkylamino; or R₇ and R₈, together with the Natom to which they are attached, form a 4 to 11-memberedheterocycloalkyl ring having 0 to 2 additional heteroatoms, or R₇ andR₈, together with the C atom to which they are attached, form C₃-C₈cycloalkyl or a 4 to 11-membered heterocycloalkyl ring having 1 to 3heteroatoms, and each of the 4 to 11-membered heterocycloalkyl rings orC₃-C₈ cycloalkyl formed by R₇ and R₈ is optionally substituted with oneor more -Q₆-T₆, wherein Q₆ is a bond, C(O), C(O)NR_(m), NR_(m)C(O),S(O)₂, or C₁-C₃ alkyl linker, R_(m) being H or C₁-C₆ alkyl, and T₆ is H,halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to7-membered heterocycloalkyl, 5- or 6-membered heteroaryl, orS(O)_(p)R_(p) in which p is 0, 1, or 2 and R_(p) is C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-memberedheterocycloalkyl, or 5- or 6-membered heteroaryl, and T₆ is optionallysubstituted with one or more substituents selected from the groupconsisting of halo, C₁-C₆ alkyl, hydroxyl, cyano, C₁-C₆ alkoxyl, amino,mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀aryl, 4 to 7-membered heterocycloalkyl, and 5- or 6-membered heteroarylexcept when T₆ is H, halo, hydroxyl, or cyano; or -Q₆-T₆ is oxo; and R₁₄is absent, H, or C₁-C₆ alkyl optionally substituted with one or moresubstituents selected from the group consisting of halo, hydroxyl, COOH,C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-memberedheterocycloalkyl, and 5- or 6-membered heteroaryl.
 3. The method ofclaim 1, wherein the compound is of Formula (Ia):


4. The method of claim 1, wherein R₆ is phenyl substituted with one ormore -Q₂-T₂.
 5. The method of claim 1, wherein R₆ is 5- or 6-memberedheteroaryl containing 1-3 additional heteroatoms selected from N, O, andS and optionally substituted with one or more -Q₂-T₂.
 6. The method ofclaim 5, wherein R₆ is pyridinyl, pyrazolyl, pyrimidinyl, quinolinyl,tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, furyl, orthienyl, each of which is optionally substituted with one or more-Q₂-T₂.
 7. The method of claim 1, wherein T₂ is —NR_(a)R_(b) or—C(O)NR_(a)R_(b), in which each of R_(a) and R_(b), independently is Hor C₁-C₆ alkyl, or R_(a) and R_(b), together with the N atom to whichthey are attached, form a 4 to 12-membered heterocycloalkyl ring having0 or 1 additional heteroatom, the C₁-C₆ alkyl and the 4 to 12-memberedheterocycloalkyl ring being optionally substituted with one or more-Q₃-T₃.
 8. The method of claim 1, wherein Q₂ is C₁-C₃ alkyl linkeroptionally substituted with halo or hydroxyl.
 9. The method of claim 1,wherein R₇ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl or 4 to 12-memberedheterocycloalkyl, each optionally substituted with one or more -Q₅-T₅.10. The method of claim 9, wherein R₇ is piperidinyl, tetrahydropyran,tetrahydro-2H-thiopyranyl, cyclopentyl, or cyclohexyl, each optionallysubstituted with one or more -Q₅-T₅.
 11. The method of claim 1, whereinone or more -Q₅-T₅ are oxo; or when Q₅ is a bond, T₅ is amino,mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₁-C₆ alkyl, C₃-C₈cycloalkyl, or 4 to 12-membered heterocycloalkyl; or when Q₅ is CO,S(O)₂, or NHC(O), T₅ is C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈ cycloalkyl, or4 to 12-membered heterocycloalkyl; or when Q₅ is C₁-C₃ alkyl linker, T₅is H, C₆-C₁₀ aryl, C₃-C₈₈ cycloalkyl, 4 to 12-membered heterocycloalkyl,or S(O)_(q)R_(q).
 12. The method of claim 1, wherein R₁₁ is H.
 13. Themethod of claim 1, wherein each of R₂ and R₄, independently is H orC₁-C₆ alkyl optionally substituted with amino, mono-C₁-C₆ alkylamino,di-C₁-C₆ alkylamino, or C₆-C₁₀ aryl.
 14. The method of claim 1, whereinR₁ is H.
 15. The method of claim 1, wherein R₁₂ is H, methyl, ethyl,ethenyl, or halo.
 16. The method of claim 1, wherein R₈ is H, methyl, orethyl.
 17. The method of claim 1, wherein Z is NR₇R₈ or CR₇R₈R₁₄ whereinR₇ and R₈, together with the atom to which they are attached, form aring selected from the group consisting of piperidinyl, morpholinyl,piperazinyl, and cyclohexenyl, each optionally substituted with one-Q₆-T₆.
 18. The method of claim 1, wherein R₁₃ is H or methyl.
 19. Themethod of claim 1, wherein the compound is of Formula (Ie):


20. The method of claim 1, wherein the compound is of Formula (Ig):

wherein R₂, R₄ and R₁₂ are each, independently C₁₋₆ alkyl.
 21. Themethod of claim 20, wherein R₆ is C₆-C₁₀ aryl or 5- or 6-memberedheteroaryl, each of which is optionally, independently substituted withone or more -Q₂-T₂, wherein Q₂ is a bond or C₁-C₃ alkyl linker, and T₂is H, halo, cyano, —OR_(a), —NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻,—C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a), —S(O)₂R_(a), or R_(S2), in whicheach of R_(a) and R_(b), independently is H or R_(S3), each of R_(S2)and R_(S3), independently, is C₁-C₆ alkyl, or R_(a) and R_(b), togetherwith the N atom to which they are attached, form a 4 to 7-memberedheterocycloalkyl ring having 0 or 1 additional heteroatom, and each ofR_(S2), R_(S3), and the 4 to 7-membered heterocycloalkyl ring formed byR_(a) and R_(b), is optionally, independently substituted with one ormore -Q₃-T₃, wherein Q₃ is a bond or C₁-C₃ alkyl linker and T₃ isselected from the group consisting of halo, C₁-C₆ alkyl, 4 to 7-memberedheterocycloalkyl, OR_(d), —S(O)₂R_(d), and —NR_(d)R_(e), each of R_(d)and R_(e) independently being H or C₁-C₆ alkyl, or -Q₃-T₃ is oxo; or anytwo neighboring -Q₂-T₂, together with the atoms to which they areattached form a 5- or 6-membered ring optionally containing 1-4heteroatoms selected from N, O and S.
 22. The method of claim 2, whereinthe compound is of Formula (II):

wherein Q₂ is a bond or methyl linker, T₂ is H, halo, —OR_(a),—NR_(a)R_(b), —(NR_(a)R_(b)R_(c))⁺A⁻, or —S(O)₂NR_(a)R_(b), R₇ ispiperidinyl, tetrahydropyran, cyclopentyl, or cyclohexyl, eachoptionally substituted with one -Q₅-T₅ and R₈ is ethyl.
 23. The methodof claim 20, wherein the compound is of Formula (IIa):


24. The method of claim 23, wherein R_(a) and R_(b), together with the Natom to which they are attached, form a 4 to 7-membered heterocycloalkylring having 0 or 1 additional heteroatoms to the N atom and the ring isoptionally substituted with one or more -Q₃-T₃, wherein theheterocycloalkyl is azetidinyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,or morpholinyl.
 25. The method of claim 24, wherein R₇ is C₃-C₈cycloalkyl or 4 to 7-membered heterocycloalkyl, each optionallysubstituted with one or more -Q₅-T₅.
 26. The method of claim 25, whereinR₇ is piperidinyl, tetrahydropyran, tetrahydro-2H-thiopyranyl,cyclopentyl, cyclohexyl, pyrrolidinyl, or cycloheptyl, each optionallysubstituted with one or more -Q₅-T₅.
 27. The method of claim 26, R₈ is Hor C₁-C₆ alkyl which is optionally substituted with one or moresubstituents selected from the group consisting of halo, hydroxyl, COOH,C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,and di-C₁-C₆ alkylamino.
 28. The method of claim 1, wherein the compoundis selected from Compounds 13, 1, 2, 11, 12, 17, 20, 21, 36, 42, 43, 44,59, 65, 67, 68, 69, 73, 75, 76, 78, 80, 82, 83, 87, 88, 89, 90, 91, 94,97, 103, 105, 138 and 141, and their pharmaceutically acceptable saltsthereof.
 29. A method of treating soft tissue sarcoma, comprisingadministering to a

subject in need thereof a therapeutically effective amount of or apharmaceutically acceptable salt thereof.